Applications of Nanoparticle Tracking Analysis in Nanomedicine

Nanoparticles are increasingly being used in a wide range of sectors. This article evaluates particular mechanisms through which nanoparticles are uniquely developed and formulated. It also discusses the important role of nanoparticle tracking analysis (NTA) in the field of nanomedicine.

Banerjee et al. (2010) studied magnetic nanoparticles and their role and applications in nanomedicine; Villaverde (2011) assessed the emergence of nanoparticles in both medicine and translational science; and Stanishevsky et al. (2011) reviewed the effect and biomedical applications of nanostructured carbiobeads, which are a subset of nanoparticles.

While analyzing the issues and opportunities in the development of nanomedicines, Wei et al. (2012) identified a number of requirements, including general and powerful techniques for the precise characterization of shape, size, and composition of nanoparticles, and also particle engineering to ensure adequate stability during storage and to sustain low levels of nonspecific cytotoxicity.

Both DLS and NTA techniques were compared when performing size analysis of nanocarriers that contained 50:50 poly(lactic-co-glycolic acid) (PLGA), and commercial liposomes as well as trimethyl chitosan. The results showed that DLS often fails to report precise data, except in the case of a monodisperse samples.

Recently, Bai et al. (2012) showed that uniform submicron particles can actually affect the growth of larger sized articles following long-standing storage in a temperature-dependent way. This implied product stability at which point Interferon-beta-1a was stressed thermally at 50°C temperature for a period of 6 hours and then subsequently defined by means of circular dichroism (CD) spectroscopy, a microflow digital imaging (MFI), and NTA techniques.

The role, effect, and characterization of the NTA technique was recently investigated, after an early review of NTA as an evolving method (Filipe et al., 2010). Cho et al. (2013) also talked about the emerging technologies and challenges related to nanoparticle characterization, as these technologies have been shown to be a promising tool towards improved drug delivery and diagnosis.

This article gives a critical review of in vivo and in vitro methods that are presently being employed to assess nanoparticles, and also discusses new modes and methods, including NTA, which may be complementarily utilized.

Liposomes, Microvesicles and Micelles

Sorrell and Lyon (2008) analyzed the deformation-controlled arrangement of binary microgel thin films following a prior work, in which NTA was used for the characterization of casein micelles and in the dispersion of poly(3,4-ethylenedioxythiophene) in organic liquids.

Micellar systems have low light scattering properties, and therefore cannot be easily detected on an individual basis. Despite this fact, NTA has been effectively utilized to define these types of structures, and specifically in the development of drug delivery micellar formulations for the encapsulation of mithramycin and for the regulated release of doxorubicin that is covalently entrapped.

This work showed that when compared to batch systems, microfluidics represents a robust technique for microfluidic nanoprecipitation-based formulation of drug-loaded polymeric micelles, as it facilitated improved control, homogeneity, and reproducibility of the size properties of the produced micelles.

Morton et al. (2012) applied EM, DLS, and NTA techniques to define the product's monodispersity in order to enhance the homogeneity of nano-sized lipid vesicles as drug delivery systems. NTA was also used by Wrenn et al. (2012) to find out the number of liposomes and their related diameter under an ultrasound. This was done to differentiate the mechanisms as well as to measure the contributions of liposome destruction against nanoparticle diffusion via the bilayer.

It was observed that the overall number of liposomes reduced with ultrasound exposure time; however, the most marked decrease was seen in the initial four minutes of ultrasound exposure. This indicates that some vesicle destruction does takes place and this was found to be consistent with the earlier work. NTA was also used by Brinkhuis et al. (2012) to determine the zeta potential of polymersomes and to ultimately demonstrate that size indeed affects the pharmacokinetics. As a result, long circulating preparations must be less than 100nm.

Encapsulation

In yet another study, curcumin-loaded lipid-core nanocapsules (C-LNC) were developed to improve the antiglioma activity of this type of polyphenol. Here, NTA was used to view the C-LNC, and the data thus acquired indicated that the curcumin nanoencapsulation in LNC is a key approach to enhance its pharmacological efficacy in glioma treatment.

In a case study using BaTiO₃, the surface functionalization of the metal oxide nanoparticles was studied using biologically active molecules comprising phosphonate moieties. Through a variety of methods such as energy dispersive spectroscopy/scanning electron microscopy, IR and NMR spectroscopy, pH-metric titration, ζ potential, DLA, thermogravimetric analysis, NTA, and radiometric measurements, it was observed that using amino phosphonic acids as surface ligands rendered significant solution stability to the nanoparticles under an aqueous medium and at neutral pH conditions, and most importantly in the presence of electrolytes. These results offer many potential applications for nanoparticle dispersions in the fields of nanomagnetism and nano-optics.

While designing and developing drug delivery systems that are capable of buccal delivery, Mazzarino and her team devised a chitosan-coated nanoparticles that were loaded with curcumin for mucoadhesive applications. At varied concentrations and scattering angles, DLS studies revealed that the nanoparticles are indeed monodisperse. NTA was applied to assess the nanoparticle systems, and the results were found to be consistent with those acquired by the DLS method. When coated with chitosan, the nanoparticles were effectively able to interact with mucin, thus suggesting their utility for mucoadhesive applications.

Delivery and Controlled Release

One of the major challenges and opportunities in nanomedicine is to design nano- and micro-particles that can release drug cargos in a certain location at a particular time. A better understanding of the number concentration, size, and size distribution profile is important in the design and development of such systems. In this regard, NTA has been shown to be useful in providing this data at all stages of the production process. Therefore, the behavior of complicated multifunctional structures in biological environments, core particle size, and the efficiency of addition of functionalized coatings has been extensively studied. Some examples of NTA applications are discussed in the following sections.

Previous studies suggested the potential applications of NTA in the analysis of complex and multifunctional nanoparticles, and the subsequent work dealt with different types of particles and applications. Park et al. (2011) addressed nanoparticle functionalization in their work, which was focused on the improvement of surface ligand display on PLGA nanoparticles having amphiphilic ligand conjugates, and Kusters et al. (2011) reported their study on the immobilization of biological membranes in hydrogels. In both these examples, NTA was employed to track particle size during all stages of development.

Recently, advanced applications and structures have been developed, wherein NTA was applied to establish their structure and thus improve the production method. Chang et al. (2013) described an aggregation-induced photodynamic therapy improvement, which was built on nonlinear and linear excited FRET of fluorescent organic nanoparticles, demonstrating that a binary molecule can assemble on its own to form fluorescent organic nanoparticles. Here, NTA was used to determine nanoparticle sizes as well as the sizing partition curves.

According to Reis et al. (2013), although formulation for protein and peptide delivery via the oral route has always remained a key strategy with the development of biotechnology, ineffective absorption, enzymatic degradation, and stability are some of the common issues that are seen in traditional dosage forms. These observations highlight the requirement for novel drug-delivery methods that could overcome these restrictions, and thus improve oral drug delivery.

Design and Formulation

NTA was used by Khan et al. (2011) to determine particle sizes and concentrations and to reveal the effects of His-affinity tags on levels of protein expression, secondary structure, solubility, aggregation, thermal denaturation, as well as the effect on cellular and humoral immune responses in mice. These outcomes indicated that the utility of protein tags may be offset by their probable effect on function and structure, thus underscoring the need to exercise caution while using them.

Sunshine et al. (2012) used NTA to characterize the samples, and demonstrated that both uptake and transfection with polymeric nanoparticles rely on polymer end-group structure, but is mostly independent of chemical and physical properties of nanoparticles.

Liling (2008) had used NTA in his analysis of bio-responsive peptide-inorganic nanomaterials, and Troiber et al. (2012) reviewed NTA among three other sizing methods during the comparison of four different types of particle sizing techniques for the characterization of siRNA polyplex.

Zhuang et al. (2013) used NTA to assess macromolecules and their corresponding assemblies to define micelles under fluorescent as well as light scatter modes so as to elucidate the origin of the mechanisms of stimuli responsiveness. All these data can provide a guideline to develop futuristic multi-stimuli responsive materials.

Conclusion

Nanoparticles are playing a major role in the emerging field of nanomedicine. Since the size, number concentration, and size distribution profile is key to the development of target-specific drug delivery systems, NTA, in this regard, has been shown to be extremely useful during all stages of the manufacturing process.

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