Effective delivery of the anti-mycobacterial peptide NZX in mesoporous silica nanoparticles.

BACKGROUND: Intracellular delivery of antimicrobial agents by nanoparticles, such as mesoporous silica particles (MSPs), offers an interesting strategy to treat intracellular infections. In tuberculosis (TB), Mycobacterium tuberculosis avoids components of the immune system by residing primarily inside alveolar macrophages, which are the desired target for TB therapy. METHODS AND FINDINGS: We have previously identified a peptide, called NZX, capable of inhibiting both clinical and multi-drug resistant strains of M. tuberculosis at therapeutic concentrations. In this study we analysed the potential of MSPs containing NZX for the treatment of tuberculosis. The MSPs released functional NZX gradually into simulated lung fluid and the peptide filled MSPs were easily taken up by primary macrophages. In an intracellular infection model, the peptide containing particles showed increased mycobacterial killing compared to free peptide. The therapeutic potential of peptide containing MSPs was investigated in a murine infection model, showing that MSPs preserved the effect to eliminate M. tuberculosis in vivo. CONCLUSIONS: In this study we found that loading the antimicrobial peptide NZX into MSPs increased the inhibition of intracellular mycobacteria in primary macrophages and preserved the ability to eliminate M. tuberculosis in vivo in a murine model. Our studies provide evidence for the feasibility of using MSPs for treatment of tuberculosis.

Quantitative 19F MRI of perfluoro-15-crown-5-ether using uniformity correction of the spin excitation and signal reception.

OBJECTIVES: A common limitation of all 1H contrast agents is that they only allow indirect visualization through modification of the intrinsic properties of the tissue, making quantification of this effect challenging. 19F compounds, on the contrary, are measured directly, without any background signal. There is a linear relationship between the amount of 19F spins and the intensity of the signal. However, non-uniformity of the radiofrequency field may lead to errors in the quantified 19F signal and should be carefully addressed for any quantitative imaging. MATERIALS AND METHODS: Adaptation of the previously introduced [Formula: see text] mapping technique to the problem of quantifying the 19F signal from perfluoro-15-crown-5-ether (PFCE) is proposed in this work. Initial evaluation of the proposed technique simultaneously accounting for transmit [Formula: see text] and receive [Formula: see text] field inhomogeneities is performed in a PFCE phantom. As a proof of concept, in vivo quantification of the 19F signal is performed in a murine model after application of custom-designed hollow mesoporous silica spheres (HMSS) loaded with PFCE. RESULTS: A phantom experiment clearly shows that only compensation for both transmit and receive characteristics outperforms inaccurate quantification based on the non- or partly-corrected signal intensities. Furthermore, an optimized protocol is proposed for in vivo application. CONCLUSION: The proposed [Formula: see text]/[Formula: see text] mapping technique represents a simple to implement and easy-to-use solution for quantification of the 19F signal from PFCE in the presence of B1-field inhomogeneities.

Cargo-influences on the biodistribution of hollow mesoporous silica nanoparticles as studied by quantitative 19F-magnetic resonance imaging.

HYPOTHESIS: Biodistribution is a key issue when it comes to medical applications of nanomaterials. Hollow mesoporous silica nanoparticles (HMSNs) loaded with fluorine compounds can be applied as positive magnetic resonance imaging (MRI) contrast agents (CAs). These CAs exhibit an unusual biodistribution which is influenced by the cargo and which could be linked to their serum protein adsorption behaviour. EXPERIMENTS: HMSNs were post-synthetically loaded with perfluoro-15-crown-5-ether (PFCE). The 19F signal was quantified with MRI in a murine model. Furthermore protein adsorption tests were performed in full serum. FINDINGS: Quantitative analysis of the 19F-signal revealed that the particles were exclusively accumulating in the liver 24h post-injection, and no accumulation in other reticuloendothelial system (RES) organs like spleen or lung was observed. The protein corona around non-loaded and loaded particles was therefore analysed, and more proteins adsorbed on PFCE-loaded particles as compared to the bare particles, and importantly, the amount of apolipoproteins A-1 and A-2, was clearly elevated for the PFCE-loaded particles. The results underline that the type of cargo may have major influences on the biodistribution of mesoporous silica drug vectors.

Membrane interactions of mesoporous silica nanoparticles as carriers of antimicrobial peptides.

Membrane interactions are critical for the successful use of mesoporous silica nanoparticles as delivery systems for antimicrobial peptides (AMPs). In order to elucidate these, we here investigate effects of nanoparticle charge and porosity on AMP loading and release, as well as consequences of this for membrane interactions and antimicrobial effects. Anionic mesoporous silica particles were found to incorporate considerable amounts of the cationic AMP LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES (LL-37), whereas loading is much lower for non-porous or positively charged silica nanoparticles. Due to preferential pore localization, anionic mesoporous particles, but not the other particles, protect LL-37 from degradation by infection-related proteases. For anionic mesoporous nanoparticles, membrane disruption is mediated almost exclusively by peptide release. In contrast, non-porous silica particles build up a resilient LL-37 surface coating due to their higher negative surface charge, and display largely particle-mediated membrane interactions and antimicrobial effects. For positively charged mesoporous silica nanoparticles, LL-37 incorporation promotes the membrane binding and disruption displayed by the particles in the absence of peptide, but also causes toxicity against human erythrocytes. Thus, the use of mesoporous silica nanoparticles as AMP delivery systems requires consideration of membrane interactions and selectivity of both free peptide and the peptide-loaded nanoparticles, the latter critically dependent on nanoparticle properties.