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壳聚糖包被的尼欧索米与肠道细胞单层相互作用的表征和实时无标记评估。

Characterization and Real-Time Label-Free Assessment of the Interaction of Chitosan-Coated Niosomes with Intestinal Cellular Monolayers.

机构信息

Drug Research Program, Division of Pharmaceutical Chemistry and Technology, Faculty of Pharmacy, University of Helsinki, Helsinki 00014, Finland.

Department of Pharmacy, University of Chieti - Pescara "G. d'Annunzio", Chieti 66100, Italy.

出版信息

Langmuir. 2023 Jun 13;39(23):8255-8266. doi: 10.1021/acs.langmuir.3c00728. Epub 2023 Jun 2.

Abstract

cell-based characterization methods of nanoparticles are generally static and require the use of secondary analysis techniques and labeling agents. In this study, bare niosomes and chitosan-coated niosomes (chitosomes) and their interactions with intestinal cells are studied under dynamic conditions and without fluorescent probes, using surface plasmon resonance (SPR)-based cell sensing. Niosomes and chitosomes were synthesized by using Tween 20 and cholesterol in a 15 mM:15 mM ratio and then characterized by dynamic light scattering (DLS). DLS analysis demonstrated that bare niosomes had average sizes of ∼125 nm, polydispersity index (PDI) below 0.2, and a negative zeta (ζ)-potential of -35.6 mV. In turn, chitosomes had increased sizes up to ∼180 nm, with a PDI of 0.2-0.3 and a highly positive ζ-potential of +57.9 mV. The viability of HT29-MTX, Caco-2, and Caco-2/HT29-MTX cocultured cells showed that both niosomes and chitosomes are cytocompatible up to concentrations of 31.6 μg/mL for at least 240 min. SPR analysis demonstrated that chitosomes interact more efficiently with HT29-MTX, Caco-2, and Caco-2/HT29-MTX cocultures compared to bare niosomes. The resulting SPR measurements were further supported by confocal microscopy and flow cytometry studies, which demonstrated that this method is a useful complementary or even alternative tool to directly characterize the interactions between niosomes and cell models in label-free and real-time conditions.

摘要

基于细胞的纳米粒子特性分析方法通常是静态的,需要使用二次分析技术和标记试剂。在本研究中,我们使用基于表面等离子体共振(SPR)的细胞传感技术,在动态条件下且不使用荧光探针的情况下,研究了裸囊泡和壳聚糖包覆囊泡(壳囊泡)及其与肠道细胞的相互作用。采用 Tween 20 和胆固醇以 15mM:15mM 的比例合成了囊泡和壳囊泡,并通过动态光散射(DLS)进行了表征。DLS 分析表明,裸囊泡的平均粒径约为 125nm,多分散指数(PDI)低于 0.2,且ζ-电位为负(-35.6mV)。相反,壳囊泡的粒径增大到约 180nm,PDI 为 0.2-0.3,ζ-电位为正(+57.9mV)。HT29-MTX、Caco-2 和共培养的 Caco-2/HT29-MTX 细胞的活力表明,裸囊泡和壳囊泡在至少 240 分钟内,在 31.6μg/mL 浓度下都是细胞相容的。SPR 分析表明,与裸囊泡相比,壳囊泡与 HT29-MTX、Caco-2 和共培养的 Caco-2/HT29-MTX 细胞相互作用更有效。共聚焦显微镜和流式细胞术研究进一步支持了这些 SPR 测量结果,表明该方法是一种有用的补充方法,甚至是一种替代方法,可以在无标记和实时条件下直接分析囊泡与细胞模型之间的相互作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/64d1/10269435/5c2e40bb11fe/la3c00728_0001.jpg

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