The different ways to chitosan/hyaluronic acid nanoparticles: templated vs direct complexation. Influence of particle preparation on morphology, cell uptake and silencing efficiency

Abstract

This study is about linking preparative processes of nanoparticles with the morphology of the nanoparticles and with their efficiency in delivering payloads intracellularly. The nanoparticles are composed of hyaluronic acid (HA) and chitosan; the former can address a nanoparticle to cell surface receptors such as CD44, the second allows both for entrapment of nucleic acids and for an endosomolytic activity that facilitates their liberation in the cytoplasm. Here, we have systematically compared nanoparticles prepared either A) through a two-step process based on intermediate (template) particles produced via ionotropic gelation of chitosan with triphosphate (TPP), which are then incubated with HA, or B) through direct polyelectrolyte complexation of chitosan and HA. Here we demonstrate that HA is capable to quantitatively replace TPP in the template process and significant aggregation takes place during the TPP–HA exchange. The templated chitosan/HA nanoparticles therefore have a mildly larger size (measured by dynamic light scattering alone or by field flow fractionation coupled to static or dynamic light scattering), and above all a higher aspect ratio (R g/R H) and a lower fractal dimension. We then compared the kinetics of uptake and the (antiluciferase) siRNA delivery performance in murine RAW 264.7 macrophages and in human HCT-116 colorectal tumor cells. The preparative method (and therefore the internal particle morphology) had little effect on the uptake kinetics and no statistically relevant influence on silencing (templated particles often showing a lower silencing). Cell-specific factors, on the contrary, overwhelmingly determined the efficacy of the carriers, with, e.g., those containing low-MW chitosan performing better in macrophages and those with high-MW chitosan in HCT-116.

Attenuation of Radiation-Induced Lung Injury by Hyaluronic Acid Nanoparticles

“…In the present study, we observed that the size of HANPs, but also the preincubation period and the IR insult, contributed to the final effect on cell viability (Choi et al, 2010). Consistent with our results, Gennari et al, (2019) confirmed a size-dependent effect of chitosan/HA nanoparticles on cell uptake and silencing efficiency in macrophages (Gennari et al, 2019). Recent studies of HANPs' biodistribution and stability in organisms have demonstrated the most intensive tissues accumulation to be in liver, tumor, and lungs.…”

Asymmetric flow field-flow fractionation as a multifunctional technique for the characterization of polymeric nanocarriers

“…This kind of analysis can turn useful during carrier formulation to detect the formation of aggregates. As an example, a reported work [94] aimed to compare particles of chitosan and hyaluronate formulated in two alternative ways, by simply mixing the two polymers or by adding TPP as a template to promote the gelification of chitosan before adding hyaluronic acid. While both kinds of particles displayed values of D m > 2, typical for a spherical shape, those obtained by template addition were significantly smaller, suggesting that the hyaluronate forms a less dense shell around the chitosan core.…”

Formulation development and evaluation of lidocaine hydrochloride loaded in chitosan-pectin-hyaluronic acid polyelectrolyte complex for dry socket treatment

“…The particle size and zeta potential increased as the concentration of LH and HA increased. The HA concentration had a greater effect on size than LH due to HA’s negative charge ( Table 1 ), allowing it to form cross-links and possibly coat the particle ( Gennari et al, 2019 ).…”