Cellular Uptake and Cytotoxicity of β-Lactoglobulin Nanoparticles: The Effects of Particle Size and Surface Charge

Abstract

It is necessary to understand the cellular uptake and cytotoxicity of food-grade delivery systems, such as β-lactoglobulin (β-lg) nanoparticles, for the application of bioactive compounds to functional foods. The objectives of this study were to investigate the relationships between the physicochemical properties of β-lg nanoparticles, such as particle size and zeta-potential value, and their cellular uptakes and cytotoxicity in Caco-2 cells. Physicochemical properties of β-lg nanoparticles were evaluated using particle size analyzer. Flow cytometry and confocal laser scanning microscopy were used to investigate cellular uptake and cytotoxicity of β-lg nanoparticles. The β-lg nanoparticles with various particle sizes (98 to 192 nm) and zeta-potential values (−14.8 to −17.6 mV) were successfully formed. A decrease in heating temperature from 70°C to 60°C resulted in a decrease in the particle size and an increase in the zeta-potential value of β-lg nanoparticles. Non-cytotoxicity was observed in Caco-2 cells treated with β-lg nanoparticles. There was an increase in cellular uptake of β-lg nanoparticles with a decrease in particle size and an increase in zeta-potential value. Cellular uptake β-lg nanoparticles was negatively correlated with particle size and positively correlated with zeta-potential value. Therefore, these results suggest that the particle size and zeta-potential value of β-lg nanoparticles play an important role in the cellular uptake. The β-lg nanoparticles can be used as a delivery system in foods due to its high cellular uptake and non-cytotoxicity.

Untitled

“…As the chitosan coated (M2 and R2) formulations (22.6 ± 0.34 mV, 23.7 ± 0.03mV) have more zeta potential than uncoated (M1 and R1) formulations (-7.68 ± 0.02 mV, -6.51 ± 0.03 mV) so the % inhibition of the A549 and H1299 cells was found more in these formulations. Higher value of positive zeta potential facilitates cytotoxicity in cancer cells due to stronger interaction with tumor cell membrane 28,29 . He et al, found that an increase in zeta-potential values of carboxymethyl chitosan grafted methyl methacrylate nanoparticles resulted in a significant rise in the cellular uptake of nanoparticles in murine macrophage cells 29 .…”

“…The particle size of chitosan coated nanoliposomes appeared to be the controlling factor affecting the cytotoxicity of the batch M2. A decrease in the size of nanoliposomes may lead to an increase in the interactions and binding with cell membranes, which can result in enhanced cytotoxicity of chitosan coated nanoliposomes 28,30,31 . Zhang et al, reported that drug uptake in cancer cells reliant on the particle size.…”

“…Thus, cytotoxic activity of chitosan coated nanoliposomes increased by using nanoliposomes with smaller particle size 32 . Ha et al, demonstrated that the size and surface charge of β-lg nanoparticles were play an important role in the cellular uptake of β-lg nanoparticles in Caco-2 cells 28 .…”

Smart multifunctional nanoparticles in nanomedicine

“…Nanoparticle cell interactions can be remarkably different from that of small or large sized particles at nanoscale [24]. As nanoparticles with a smaller size have larger surface areas available to adhere and interact with cell membranes, the decrease of nanoparticle size may lead to an increase in mobility and interaction with cell membranes, which can result in enhanced cellular uptake of nanoparticles [25]. Besides, because of their small size, nanoparticles are often not recognized as a foreign agent by macrophages and consequently do not enter macrophages through membrane pores to be led to the digestion apparatus for such microparticles, the reticuloendothelial system [26].…”

Synthesis and characterization of calcium-induced peanut protein isolate nanoparticles

“…8 There are lots of different kinds of protein nanoparticles reported, such as bovine serum albumin, 9,10 casein, 11,12 collagen, 13 b-lacto globulin 14,15 and other animal proteins. 16 However, reports about the application of plant protein in the preparation of nanoparticles as the matrix material are relatively few.…”