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基于聚乳酸与维生素E的罗非鱼疫苗递送系统装置植入:生物相容性和生物降解性研究

Device implant based on poly (lactic acid) with vitamin E for vaccine delivery system in Tilapia: Study for biocompatibility and biodegradation.

作者信息

Conde Gabriel, Aracati Mayumi Fernanda, Rodrigues Letícia Franchin, de Oliveira Susana Luporini, da Costa Camila Carlino, Charlie-Silva Ives, Ruiz Thalles Fernando Rocha, Taboga Sebastião Roberto, Belo Marco Antonio de Andrade

机构信息

Department of Preventive Veterinary Medicine, São Paulo State University (Unesp), Rodovia de Acesso Paulo Donato Castellane s/n, Zona Rural, zipcode 14884-900, Jaboticabal, SP, Brazil.

Deparment of Pharmacology, Institute of Biomedical Science, University of São Paulo (USP). 2415 Prof. Lineu Prestes Avenue, Cidade Universitária, São Paulo SP, ZipCode:05508-000, Brazil.

出版信息

Fish Shellfish Immunol Rep. 2022 Jul 3;3:100060. doi: 10.1016/j.fsirep.2022.100060. eCollection 2022 Dec.

Abstract

The use of Poly (lactic acid) (PLA) as a slow-release vehicle for vaccines has attracted the attention of researchers, since its insertion improves the uptake of them, and reduces side effects or by stimulating recruited defense cells, assisting immunity without the need for booster vaccine doses. Seeking to develop new strategies for the administration of drugs and vaccines in aquaculture, we evaluated the biocompatibility and biodegradation of polymeric PLA devices and PLA plus vitamin E devices, implanted through subcutaneous (SC) and intraperitoneal (IP) routes in Nile tilapia. To carry out this study, 84 male tilapia (initial 243.82 ± 56.74 g; final 400.71 ± 100.54 g) were randomly distributed in 3 tanks ( = 28 fish per treatment/tank). The devices were prepared in two formulations: neat PLA (containing 100% PLA) and PLAVE (PLA plus vitamin E) implanted using a commercial AnimalTag® applicator, and non-implanted fish (control). Fish were sampled 15, 30, 60, and 120 days post-implantation (DPI). Blood analysis was used to access blood cells and blood smear for differential leucocytes count. Serum biochemistry to evaluated changes in serum proteins and glycemia. Histopathological investigation using hematoxylin-eosin (H&E) was used to assess polymer-tissue interaction. Histochemistry and immunohistochemistry was used to detection immune cells and phagocytes in capsule, and analyses of melanomacrophage centers (MMCs) to morphometric evaluation and percentage amount of melanin, hemosiderin and lipofucsin pigments. Histopathological study revealed an increase of capsular formation and inflammatory cell infiltration in PLAVE-implanted tilapia through SC route (15 DPI). Tilapia implanted with PLAVE and PLA (SC) presented mast cells and eosinophilic granular cells during 15, 30, and 60 DPI, with a decrease in these cells in the fibrous capsule around the polymer at 120 DPI. PLAVE implanted tilapia SC at 60 DPI showed significantly phagocytosis points than other groups. Phagocytic cells (F4/80+) were observed near to biopolymers in phagocytosis sites. Lipofuscin at 120 DPI in spleen melanomacrophage centers were significantly high in PLAVE implanted tilapias when compared to fish with PLA implants and control. The serum biochemical study of tilapia did not reveal changes in cytotoxicity and liver function in implanted fish. The absence of side effects in hematological and biochemical findings, including the absence of mortality after device implantation, proves its clinical safety. PLA implants in tilapia have demonstrated biocompatibility, biodegradation, clinical safety, and excellent evolution of foreign body inflammatory responses.

摘要

聚乳酸(PLA)作为疫苗缓释载体的应用已引起研究人员的关注,因为它的植入能提高疫苗的摄取,并通过刺激募集的防御细胞减少副作用,无需加强疫苗剂量即可辅助免疫。为寻求水产养殖中药物和疫苗给药的新策略,我们评估了通过皮下(SC)和腹腔内(IP)途径植入尼罗罗非鱼体内的聚乳酸(PLA)聚合物装置以及PLA加维生素E装置的生物相容性和生物降解性。为开展本研究,84条雄性罗非鱼(初始体重243.82±56.74克;最终体重400.71±100.54克)被随机分配到3个水箱中(每个处理/水箱28条鱼)。装置制备成两种配方:纯PLA(含100%PLA)和PLAVE(PLA加维生素E),使用商用AnimalTag®涂抹器植入,未植入装置的鱼作为对照。在植入后15、30、60和120天(DPI)对鱼进行采样。通过血液分析获取血细胞并制作血涂片进行白细胞分类计数。采用血清生化分析评估血清蛋白和血糖的变化。使用苏木精 - 伊红(H&E)染色进行组织病理学研究,以评估聚合物与组织的相互作用。采用组织化学和免疫组织化学检测胶囊中的免疫细胞和吞噬细胞,并对黑素巨噬细胞中心(MMCs)进行形态计量评估以及分析黑色素、含铁血黄素和脂褐素色素的含量百分比。组织病理学研究显示,通过SC途径植入PLAVE的罗非鱼在15 DPI时包膜形成增加且炎症细胞浸润增多。植入PLAVE和PLA(SC)的罗非鱼在15、30和60 DPI时出现肥大细胞和嗜酸性粒细胞,在120 DPI时聚合物周围纤维包膜中的这些细胞数量减少。在60 DPI时,通过SC途径植入PLAVE的罗非鱼的吞噬点明显多于其他组。在吞噬部位的生物聚合物附近观察到吞噬细胞(F4/80 +)。与植入PLA的鱼和对照组相比,在120 DPI时,植入PLAVE的罗非鱼脾脏黑素巨噬细胞中心的脂褐素含量显著更高。罗非鱼的血清生化研究未显示植入鱼的细胞毒性和肝功能有变化。血液学和生化检查结果中未出现副作用,包括装置植入后无死亡情况,证明了其临床安全性。PLA植入罗非鱼体内已证明具有生物相容性、生物降解性、临床安全性以及对外来物体炎症反应的良好演变。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5345/9680062/72a86d0b9f59/ga1.jpg

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