Chitosan functionalisation of gold nanoparticles encourages particle uptake and induces cytotoxicity and pro-inflammatory conditions in phagocytic cells, as well as enhancing particle interactions with serum components

Journal of Nanobiotechnology

Research

Ursula Lütz-Meindl¹  Ancuela Andosch¹  Albert Duschl²  Mirjam Zimmermann²  Matthew S. P. Boyles²  Martin Himly²  Theresa Kristl³  Christian G. Huber³  Ngoc Tran⁴  Eudald Casals⁴  Victor Puntes⁵ 

[1] Department of Cell Biology, Paris-Lodron University Salzburg, Salzburg, Austria;Department of Molecular Biology, Division of Allergy and Immunology, Paris-Lodron University Salzburg, Hellbrunnerstr. 34, 5020, Salzburg, Austria;Department of Molecular Biology, Division of Chemistry and Bioanalytics, Paris-Lodron University Salzburg, Salzburg, Austria;Institut Català de Nanotecnologia, Bellaterra, Barcelona, Spain;Institut Català de Nanotecnologia, Bellaterra, Barcelona, Spain;Institut Catala de Recerca i Estudis Avancats, Barcelona, Spain;

关键词: Charged gold nanoparticles;    Chitosan;    Exocytosis;    Pro-inflammatory responses;    Protein corona;   

DOI  :  10.1186/s12951-015-0146-9

 received in 2015-09-11, accepted in 2015-11-09,  发布年份 2015

来源: Springer

【 摘 要 】

BackgroundGold nanoparticles (AuNPs) are a popular choice for use in medical and biomedical research applications. With suitable functionalisation AuNPs can be applied in drug delivery systems, or can aid in disease diagnosis. One such functionalisation is with chitosan, which enables efficient interaction and permeation of cellular membranes, providing an effective adjuvant. As both AuNPs and chitosan have been shown to have low toxicity and high biocompatibility their proposed use in nanomedicine, either individually or combined, is expanding. However, further toxicological and immunological assessments of AuNP-chitosan conjugates are still needed. Therefore, we have evaluated how AuNP functionalisation with chitosan can affect uptake, cytotoxicity, and immunological responses within mononuclear cells, and influence the interaction of AuNPs with biomolecules within a complex biofluid. The AuNPs used were negatively charged through citrate-coating, or presented either low or high positive charge through chitosan-functionalisation. Uptake by THP-1 cells was assessed via transmission electron microscopy and electron energy loss spectroscopy, pro-inflammatory responses by ELISA and qRT-PCR, and cell death and viability via lactate dehydrogenase release and mitochondrial activity, respectively. Interactions of AuNPs with protein components of a frequently used in vitro cell culture medium supplement, foetal calf serum, were investigated using mass spectrometry.ResultsAlthough cells internalised all AuNPs, uptake rates and specific routes of intracellular trafficking were dependent upon chitosan-functionalisation. Accordingly, an enhanced immune response was found to be chitosan-functionalisation-dependent, in the form of CCL2, IL-1β, TNF-α and IL-6 secretion, and expression of IL-1β and NLRP3 mRNA. A corresponding increase in cytotoxicity was found in response to chitosan-coated AuNPs. Furthermore, chitosan-functionalisation was shown to induce an increase in unique proteins associating with these highly charged AuNPs.ConclusionsIt can be concluded that functionalisation of AuNPs with the perceived non-toxic biocompatible molecule chitosan at a high density can elicit functionalisation-dependent intracellular trafficking mechanisms and provoke strong pro-inflammatory conditions, and that a high affinity of these NP-conjugates for biomolecules may be implicit in these cellular responses.

【 授权许可】

CC BY   
© Boyles et al. 2015