In vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimus
Abstract Background To evaluate the in vivo and in vitro toxicity of a new formulation of liposome-encapsulated sirolimus (LES). Methods In vitro experiments were done using ARPE-19 and HRP cells. An MTT assay was used to determine cell metabolic activity and a TUNEL assay for detecting DNA fragment...
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doaj-f4db1abfe8c04212a491b308a8e2cd1b2021-04-02T12:18:09ZengBMCInternational Journal of Retina and Vitreous2056-99202019-09-015111010.1186/s40942-019-0186-7In vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimusMurilo Batista Abud0Ricardo Noguera Louzada1David Leonardo Cruvinel Isaac2Leonardo Gomes Souza3Ricardo Gomes dos Reis4Eliana Martins Lima5Marcos Pereira de Ávila6Federal University of GoiasFederal University of GoiasFederal University of GoiasSchool of Pharmacy, Federal University of GoiasFederal University of GoiasSchool of Pharmacy, Federal University of GoiasFederal University of GoiasAbstract Background To evaluate the in vivo and in vitro toxicity of a new formulation of liposome-encapsulated sirolimus (LES). Methods In vitro experiments were done using ARPE-19 and HRP cells. An MTT assay was used to determine cell metabolic activity and a TUNEL assay for detecting DNA fragmentation. In vivo experiments were conducted on New Zealand albino rabbits that received intravitreal injections of empty liposomes (EL) or different concentrations of LES. Histopathological and immunohistochemical analyses were performed on the rabbit’s eyes following injection. Results Eighteen eyes of nine rabbits were used. MTT assay cell viability was 95.04% in group 1 (12.5 µL/mL LES). 92.95% in group 2 (25 µL/mL LES), 91.59% in group 3 (50 µL/mL LES), 98.09% in group 4 (12.5 µL/mL EL), 95.20% on group 5 (50 µL/mL EL), 98.53% in group 6 (50 µL/mL EL), and 2.84% on group 8 (50 µL/mL DMSO). There was no statistically significant difference among groups 1 to 7 in cell viability (p = 1.0), but the comparison of all groups with group 8 was significant (p < 0.0001). The TUNEL assay comparing two groups was not statistically significant from groups 1 to 7 (p = 1.0). The difference between groups 1 to 7 and group 8 (p < 0.0001) was significant. Histopathological changes were not found in any group. No activation of Müller cells was detected. Conclusion A novel formulation of LES delivered intravitreally did not cause in vitro toxicity, as evaluated by MTT and TUNEL assays, nor in vivo toxicity as evaluated by histopathology and immunohistochemistry in rabbit eyes.http://link.springer.com/article/10.1186/s40942-019-0186-7SirolimusLESARPE-19RapamycinUveitis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Murilo Batista Abud Ricardo Noguera Louzada David Leonardo Cruvinel Isaac Leonardo Gomes Souza Ricardo Gomes dos Reis Eliana Martins Lima Marcos Pereira de Ávila |
spellingShingle |
Murilo Batista Abud Ricardo Noguera Louzada David Leonardo Cruvinel Isaac Leonardo Gomes Souza Ricardo Gomes dos Reis Eliana Martins Lima Marcos Pereira de Ávila In vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimus International Journal of Retina and Vitreous Sirolimus LES ARPE-19 Rapamycin Uveitis |
author_facet |
Murilo Batista Abud Ricardo Noguera Louzada David Leonardo Cruvinel Isaac Leonardo Gomes Souza Ricardo Gomes dos Reis Eliana Martins Lima Marcos Pereira de Ávila |
author_sort |
Murilo Batista Abud |
title |
In vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimus |
title_short |
In vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimus |
title_full |
In vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimus |
title_fullStr |
In vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimus |
title_full_unstemmed |
In vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimus |
title_sort |
in vivo and in vitro toxicity evaluation of liposome-encapsulated sirolimus |
publisher |
BMC |
series |
International Journal of Retina and Vitreous |
issn |
2056-9920 |
publishDate |
2019-09-01 |
description |
Abstract Background To evaluate the in vivo and in vitro toxicity of a new formulation of liposome-encapsulated sirolimus (LES). Methods In vitro experiments were done using ARPE-19 and HRP cells. An MTT assay was used to determine cell metabolic activity and a TUNEL assay for detecting DNA fragmentation. In vivo experiments were conducted on New Zealand albino rabbits that received intravitreal injections of empty liposomes (EL) or different concentrations of LES. Histopathological and immunohistochemical analyses were performed on the rabbit’s eyes following injection. Results Eighteen eyes of nine rabbits were used. MTT assay cell viability was 95.04% in group 1 (12.5 µL/mL LES). 92.95% in group 2 (25 µL/mL LES), 91.59% in group 3 (50 µL/mL LES), 98.09% in group 4 (12.5 µL/mL EL), 95.20% on group 5 (50 µL/mL EL), 98.53% in group 6 (50 µL/mL EL), and 2.84% on group 8 (50 µL/mL DMSO). There was no statistically significant difference among groups 1 to 7 in cell viability (p = 1.0), but the comparison of all groups with group 8 was significant (p < 0.0001). The TUNEL assay comparing two groups was not statistically significant from groups 1 to 7 (p = 1.0). The difference between groups 1 to 7 and group 8 (p < 0.0001) was significant. Histopathological changes were not found in any group. No activation of Müller cells was detected. Conclusion A novel formulation of LES delivered intravitreally did not cause in vitro toxicity, as evaluated by MTT and TUNEL assays, nor in vivo toxicity as evaluated by histopathology and immunohistochemistry in rabbit eyes. |
topic |
Sirolimus LES ARPE-19 Rapamycin Uveitis |
url |
http://link.springer.com/article/10.1186/s40942-019-0186-7 |
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