Glucose-sensitive liposomes incorporating hydrophobically modified glucose oxidase.

Glucose-sensitive liposomes were prepared by incorporating hydrophobically modified glucose oxidase (EC 1.1.3.4.) into the liposomal bilayer of dioleoylphosphatidylethanolamine and cholesteryl hemisuccinate. For the release test, calcein, a fluorescence marker, was entrapped in the liposomes. The liposomes were stable under neutral conditions in terms of calcein release but an extensive release was observed under acidic conditions. In the experiment of glucose concentration-dependent calcein release, no release was observed for 180 min when the suspension of liposome was free of glucose. With a glucose concentration of 50 mg/dL, no appreciable amount of calcein was released for the first 20 min, and then the release rate was accelerated. At 200 mg/dL glucose concentration which is diagnostic and indicative for insulin-dependent diabetes, the lag time of calcein release became shorter and a faster response was obtained. When glucose concentration further increased to 400 mg/dL, the calcein release rate and the degree of release in 180 min were almost the same as the values when the glucose concentration was 200 mg/dL. The glucose concentration-dependent release is due to pH change, since the suspension of liposomes became acidic during the release experiments.

Release property of temperature-sensitive alginate beads containing poly(N-isopropylacrylamide).

The graft copolymer (APN) of alginate and poly(N-isopropylacrylamide) (PNIPAM) were synthesized and APN beads were prepared by dropping the aqueous solution of the copolymer into an aqueous solution of Ca(2+) solution. Alginate chains were employed to play a role in forming beads by electrostatic interactions with a multivalent ion, Ca(2+). Grafted PNIPAM segments were adopted to act as a valve for the pores of the beads, since they exhibit the properties of thermal contraction and expansion. The percent of release of blue dextran from APN beads was higher at 40 degrees C than at 25 degrees C. The difference in the release between two temperatures became more distinguishable when the content of PNIPAM in APN beads is higher. Below lower critical solution temperature (LCST), the expanded PNIPAM would close the pores of the beads, resulting in a lower release rate. Above LCST, the thermally contracted polymer would open the pores, resulting in a higher release rate. The percent of release from APN beads were investigated when the temperature of the release medium is altered. The release rate was relatively low at 25 degrees C. The temperature, however, changed up to 40 degrees C, a marked increase in the release rate was observed. These trends were found to be reproducible when the temperature was repeatedly altered between 25 and 40 degrees C. As a result, a stepwise response to the temperature alteration was obtained.

Enhanced antimicrobial activity of nisin-loaded liposomal nanoparticles against foodborne pathogens.

This study was designed to evaluate the prolonged antimicrobial stability of nisin-loaded liposome (LipoN) nanoparticles against Listeria monocytogenes and Staphylococcus aureus. The sizes of bare liposomes and LipoN were uniformly distributed between 114 and 125 nm. The nanoparticles were homogeneously dispersed in water with less than 0.2 of polydispersity index. The zeta potential value of LipoN was +17.1 mV due to the positive charged nisin, attaining 70% of loading efficiency. The minimum inhibitory concentration of LipoN against L. monocytogenes and S. aureus was 320 international unit/mL. The LipoN significantly enhanced the antimicrobial stability in brain heart infusion agar compared to free nisin. The numbers of L. monocytogenes and S. aureus exposed to LipoN were effectively reduced by more than 6 log colony-forming unit/mL after 48 and 72 h of incubation, respectively. These results provide useful information for the development of antimicrobial delivery system to improve food safety.

Enhancement of the skin-protective activities of Centella asiatica L. Urban by a nano-encapsulation process.

Aqueous extracts of Centella asiatica L. Urban were encapsulated by an edible biopolymer, gelatin, which has no effect on their cosmetic activities. The nanoparticles were w/o-type spherical liposomes that had an average diameter of 115.0nm. The encapsulation efficiency was estimated to be approximately 67%, which was relatively high for these aqueous extracts. The nanoparticles showed lower cytotoxicity (10%) in human skin fibroblast cells than the unencapsulated crude extract (15%) at 1.0mg/ml, this was possibly because a smaller amount of the extract was present in the nanoparticles. The nanoparticles efficiently reduced the expression of matrix metalloproteinase (MMP)-1 in UV-irradiated cells from 136.1% to 77.6% (UV-irradiated control) and inhibited hyaluronidase expression (>60%) at a concentration of 0.5mg/ml, which was higher than the levels produced by the unencapsulated crude extracts. The nanoparticles had a very high flux through mouse skin and also remained at relatively large concentrations in the derma when compared to the unencapsulated crude extracts. These results clearly indicate that the skin-protective activities of C. asiatica were significantly improved through the nano-encapsulation process. These findings also imply that a crude extract can be used and have the same efficacy as purified compounds, which should reduce the purification process and production costs.

Glucose-sensitivity of liposomes incorporating conjugates of glucose oxidase and poly(N-isopropylacrylamide-co-methacrylic acid-co-octadecylacrylate).

Liposomes, which release their contents in response to the concentration of glucose, were prepared by modifying the liposomal surface with the conjugate of poly(N-isopropylacrylamide-co-methacrylic acid-co-octadecylacrylate) (P(NIPAM-co-MAA-co-ODA)) and glucose oxidase (GOD). The maximum enzymatic activity of copolymer conjugated GOD (Polym-GOD) was observed around pH 5.0 and the value was about 40% of that of native GOD. Nine lysine residues per GOD molecule, on average, were found to be covalently attached to the copolymers. Egg phosphatidylcholine liposomes bearing Polym-GOD released their contents in response to the concentration of glucose and the sensitivity was higher than dipalmitoylphosphatidylcholine liposomes.