Application of stabilizer improves stability of nanosuspended branched-chain amino acids and anti-inflammatory effect in LPS-induced RAW 264.7 cells.

This study investigated the use of polyglyceryl esters (PGE) as stabilizer in improving stability and anti-inflammatory activity of nanosuspended branched-chain amino acids (BCAAs). BCAAs nanosuspended with stabilizer (BS) exhibited improved stability at concentration of 5% saturation level during storage as compared to BCAAs nanosuspended with aqueous solution (BA). Additionally, anti-inflammatory activity of BS was found to be greater than that of BA. Nitric oxide scavenging activity was found to be dose-dependent, with activity of BS in sodium nitroprusside system being significantly higher than that of BA (p < 0.05) at 2.5-20 mg/mL. BS also possesses greater inhibitory activity on production of pro-inflammatory factors including inducible nitric oxide synthase, cyclooxygenase-2 (COX-2), interleukin-1ß (IL-1ß), interleukin-6 (IL-6) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells through suppressed phosphorylation of p65 subunit of NF-κB at 0.5, 2, 8 mg/mL. These results suggest that PGE used as stabilizer improves solubility and biological activity of nanosuspended BCAAs.

Influence of biopolymers on the solubility of branched-chain amino acids and stability of their solutions.

This study confirmed the possibility of biopolymer-type stabilizers to increase the saturation concentration of branched-chain amino acids by preventing their crystallization/precipitation. Although microfluidization increased the initial solubility, it failed to increase the saturation concentration of the branched-chain amino acids. The saturation concentration of the branched-chain amino acids increased from 3.81% to 4.42% and 4.85% after the incorporation of food hydrocolloids and proteins, respectively. However, the branched-chain amino acids:stabilizer ratio did not affect the solubility. In the case of food hydrocolloid-based solutions, crystal formation and growth of branched-chain amino acids occurred during storage, resulting in the precipitation of branched-chain amino acid crystals. However, food proteins effectively increased the stability of the solubilized branched-chain amino acids. The improved solubility and stability of the solubilized branched-chain amino acids could be attributed to interactions between the functional groups (carboxyl, amine, sulfate, aliphatic, aromatic, etc.) of the stabilizer and the branched-chain amino acid molecules.

Nanosuspended branched chain amino acids: the influence of stabilizers on their solubility and colloidal stability.

This study examined the influence of stabilizers with different hydrophilic-lipophilic balances on the solubility of branched chain amino acids (BCAA) and colloidal stability of nanosuspended BCAAs. Initial BCAA solubility increased by homogenization as evidenced by the BCAA solubility in Tween 80-based nanosuspensions, which remained at almost 97% of their initial solubility after 20 days of storage. However, the contents of solubilized BCAAs in Span 80-based nanosuspensions decreased to approximately 85% of their initial solubility after 20 days of storage. In fact, the BCAA:Tween 80 ratio had no effect on the colloidal stability but the same variable changed according to the BCAA:Span 80 ratio. Based on this study, it can be concluded that stabilizers with a hydrophilic trait (Tween 80) could be more effective in improving BCAA solubility and the colloidal stability of nanosuspended BCAAs compared to those with a lipophilic trait (Span 80).

Anti-inflammatory and anti-genotoxic activity of branched chain amino acids (BCAA) in lipopolysaccharide (LPS) stimulated RAW 264.7 macrophages.

The purpose of this study was to evaluate the anti-inflammatory and anti-genotoxic activity of branched-chain amino acids (BCAAs) in lipopolysaccharide (LPS) stimulated RAW 264.7 macrophages. BCAAs inhibited LPS-induced NO production, with 100 mM leucine having the most pronounced effect, suppressing NO production by 81.15%. Valine and isoleucine also reduced NO production by 29.65 and 42.95%, respectively. Furthermore, BCAAs suppressed the inducible nitric oxide synthase mRNA expression. Additionally, BCAAs decreased the mRNA expression of interleukin-6 and cyclooxygenase-2 which are proinflammatory mediators. Anti-genotoxic activities of BCAAs were assessed using the alkaline comet assay and valine, isoleucine, and leucine significantly (p < 0.05) decreased tail length of DNA (damaged portion) to 254.8 ± 7.5, 235.6 ± 5.6, and 271.5 ± 19.9 µm compared than positive control H2O2 (434.3 ± 51.3 µm). These results suggest that BCAAs can be used in the pharmaceutical or functional food industries as anti-inflammatory agents or anti-cancer agents.

Influence of lysolecithin and Tween 80 on the colloidal stability of branched chain amino acids in a nanosuspension system.

This study examined the influence of stabilizers on the solubility and colloidal stability of branched chain amino acids (BCAAs) nanosuspended through high pressure homogenization at 70°C. Although homogenization increased the initial BCAA solubility, irrespective of pH (pH 3 or 6), homogenization alone was not sufficient to increase their long-term solubility. The incorporation of stabilizers into nanosuspensions increased the saturation concentration of BCAAs but the effect of stabilizers on the increase in the saturation concentration of BCAAs was more pronounced at pH 6.0. At pH 6, Tween 80 dramatically increased the colloidal stability of the BCAA nanosuspensions, independent of the BCAA:stabilizer ratio but not at pH 3. However, the effect of lysolecithin on the colloidal stability of nanosuspended BCAAs varied depending on pH and BCAA:lysolecithin ratio. In lysolecithin-related nanosuspensions, there was no clear relationship between the colloidal stability and nanosuspension conditions including pH and BCAA:lysolecithin ratio. This study could provide a useful information on stabilizer selection for the development of liquid or colloidal products with improved solubility and colloidal stability of nanosuspended BCAAs.