RESUMO
Objective: The expression of therapeutic proteins in plant oil body bioreactors has attracted much attention. But its safety is not yet clear. This article determines the risk of safety after using the drug. Methods: The oil body-linked oleosin-hEGF microgel emulsion (OBEME) was prepared by mixing the xanthan gum with suitable concentrations in an appropriate proportion. Skin irritation and sensitization reaction were investigated in rats and guinea pigs using OBEME as test article.Results: The OBEME did not produce dermal erythema/eschar or oedema responses. The dermal subacute and subchronic toxicity of OBEME were evaluated in accordance with OECD guidelines. Compared with the control group, the basic physical signs, such as weight, feed, drinking, excretion, and behaviour of experimental animals, were not abnormal. In addition, no abnormality was found in haematological parameters, biochemical indexes, relative organ weight, and histopathological observation of organs, and there was no significant difference compared with normal saline treatment group. Therefore, we conclude that OBEME has no toxic effects and is safe and reliable to be used for topical application.
Assuntos
Portadores de Fármacos/toxicidade , Fator de Crescimento Epidérmico/toxicidade , Proteínas de Plantas/toxicidade , Proteínas Recombinantes de Fusão/toxicidade , Pele/efeitos dos fármacos , Administração Cutânea , Animais , Reatores Biológicos/efeitos adversos , Carthamus tinctorius/genética , Dermatite de Contato/diagnóstico , Dermatite de Contato/etiologia , Dermatite de Contato/patologia , Portadores de Fármacos/química , Avaliação Pré-Clínica de Medicamentos , Emulsões , Fator de Crescimento Epidérmico/administração & dosagem , Fator de Crescimento Epidérmico/genética , Eritema/induzido quimicamente , Eritema/diagnóstico , Cobaias , Humanos , Gotículas Lipídicas/química , Masculino , Microgéis , Proteínas de Plantas/administração & dosagem , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Ratos , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/genética , Pele/imunologia , Pele/lesões , Pele/patologia , Testes de Toxicidade Aguda/métodos , Testes de Toxicidade Subaguda/métodos , Testes de Toxicidade Subcrônica/métodos , Cicatrização/efeitos dos fármacosRESUMO
The purpose of this study was to prepare GeXIVA[1,2] PLGA microspheres by W/O/W re-emulsification-solvent evaporation technology and to develop sustained-release formulations to meet the clinical treatment needs of chronic neuropathic pain. Through prescription optimization, the uniformity of particle size and the encapsulation efficiency is improved, so as to achieve the quality standard of the microspheres. The mechanism of trehalose improving the stability of GeXIVA[1,2] was studied and verified by molecular simulation. The results showed that when adding trehalose to W1, using the PLGA model of 75:25, PLGA concentration of 30%, PVA concentration of 1.5%, adding 1% NaCl to PVA and adding 1% NaCl to solidification water, the prepared microspheres are smooth, the particle size is about 25 µm, and the encapsulation rate reaches 90%. The results of in vitro release experiments showed that the microspheres could be released steadily for about 30 days. The microsphere samples were characterized and analyzed by molecular simulation and powder X-ray diffractometer, and the protective mechanism of trehalose on GeXIVA[1,2] was discussed. The results showed that the hydrogen bond formed between trehalose and GeXIVA[1,2] acted as a hydration film and played a certain protective role on GeXIVA[1,2]. In addition, high-viscosity trehalose can form a glass state and wrap around GeXIVA[1,2], reducing the free movement of molecules. In the microsphere system, trehalose can also avoid the influence of PLGA material on the secondary structure of GeXIVA[1,2]. In conclusion, this study is expected to provide a new therapeutic strategy for the treatment of chronic neuropathic pain.