Nanoencapsulation of Bacteriophages in Liposomes Prepared Using Microfluidic Hydrodynamic Flow Focusing.
Nanoencapsulation of Bacteriophages in Liposomes Prepared Using Microfluidic Hydrodynamic Flow Focusing.
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DOI：
10.3389/fmicb.2018.02172
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发表时间：
2018
影响因子：
5.2
通讯作者：
Malik DJ
中科院分区：
文献类型：
--
作者：
Cinquerrui S;Mancuso F;Vladisavljević GT;Bakker SE;Malik DJ
病原微生物中抗生素耐药性的增加引起了人们对人类和动物噬菌体疗法的新兴趣。利用脂质体封装的噬菌体的“特洛伊木马”方法可以促进进入被驻留在其中的细胞内病原体感染的吞噬细胞，例如治疗由结核分枝杆菌、李斯特菌、沙门氏菌和葡萄球菌引起的感染。此外，脂质体封装的噬菌体可能粘附在含有耐药细菌的粘膜上并在其内扩散，这在治疗呼吸道和胃肠道感染中是一个挑战。由于腹泻等临床症状，口服噬菌体在胃肠道中的停留时间往往较短；这可以通过脂质体的粘膜粘附来解决。在本研究中，我们评估了使用基于微流体的技术将噬菌体封装在平均尺寸在 100 至 300 nm 之间的脂质体中。对两种模型噬菌体进行封装，即大肠杆菌 T3 足病毒（大小约 65 nm）和肌病毒金黄色葡萄球菌噬菌体 K（衣壳头约 80 nm，噬菌体尾长约 200 nm）。封装的 T3 噬菌体的产量为 109 PFU/ml，而噬菌体 K 的产量则低得多，为 105 PFU/ml。大肠杆菌 T3 噬菌体的包封率受到 T3 噬菌体聚集的影响。发现金黄色葡萄球菌噬菌体 K 与脂质体脂质双层相互作用，导致大量噬菌体结合到形成的脂质体的外部，而不是被困在脂质体内部。我们能够灭活脂质体结合的金黄色葡萄球菌 K 噬菌体，同时保留封装的噬菌体的活性，以便估计大尾噬菌体的微流体封装的产量。先前发表的关于脂质体中噬菌体封装的研究可能高估了封装的有尾噬菌体的产量。这种高估可能会影响在感染部位递送的噬菌体剂量的功效。外部结合的噬菌体将在胃酸中失活，导致低剂量的噬菌体被递送到胃肠道下游的感染部位。
Increasing antibiotic resistance in pathogenic microorganisms has led to renewed interest in bacteriophage therapy in both humans and animals. A “Trojan Horse” approach utilizing liposome encapsulated phages may facilitate access to phagocytic cells infected with intracellular pathogens residing therein, e.g., to treat infections caused by Mycobacterium tuberculosis, Listeria, Salmonella, and Staphylococcus sp. Additionally, liposome encapsulated phages may adhere to and diffuse within mucosa harboring resistant bacteria which are challenges in treating respiratory and gastrointestinal infections. Orally delivered phages tend to have short residence times in the gastrointestinal tract due to clinical symptoms such as diarrhea; this may be addressed through mucoadhesion of liposomes. In the present study we have evaluated the use of a microfluidic based technique for the encapsulation of bacteriophages in liposomes having mean sizes between 100 and 300 nm. Encapsulation of two model phages was undertaken, an Escherichia coli T3 podovirus (size ~65 nm) and a myovirus Staphylococcus aureus phage K (capsid head ~80 nm and phage tail length ~200 nm). The yield of encapsulated T3 phages was 109 PFU/ml and for phage K was much lower at 105 PFU/ml. The encapsulation yield for E. coli T3 phages was affected by aggregation of T3 phages. S. aureus phage K was found to interact with the liposome lipid bilayer resulting in large numbers of phages bound to the outside of the formed liposomes instead of being trapped inside them. We were able to inactivate the liposome bound S. aureus K phages whilst retaining the activity of the encapsulated phages in order to estimate the yield of microfluidic encapsulation of large tailed phages. Previous published studies on phage encapsulation in liposomes may have overestimated the yield of encapsulated tailed phages. This overestimation may affect the efficacy of phage dose delivered at the site of infection. Externally bound phages would be inactivated in the stomach acid resulting in low doses of phages delivered at the site of infection further downstream in the gastrointestinal tract.
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DOI：
10.1016/s1473-3099(15)00350-3
发表时间：
2016-04
期刊：
The Lancet. Infectious diseases
影响因子：
--
作者：
Kelly R;Zoubiane G;Walsh D;Ward R;Goossens H
通讯作者：
Goossens H
影响因子：
15.6
作者：
Malik, Danish J.;Sokolov, Ilya J.;Kirpichnikova, Anna
通讯作者：
Kirpichnikova, Anna
影响因子：
3.7
作者：
Denou, Emmanuel;Bruttin, Anne;Zuber, Sophie
通讯作者：
Zuber, Sophie
影响因子：
15
作者：
Jahn, A;Vreeland, WN;Locascio, LE
通讯作者：
Locascio, LE
影响因子：
2.7
作者：
Chadha, Parul;Katare, Om Prakash;Chhibber, Sanjay
通讯作者：
Chhibber, Sanjay