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. 2024 Oct 10;13(10):1218.
doi: 10.3390/antiox13101218.

The Effect of Multilayer Nanoemulsion on the In Vitro Digestion and Antioxidant Activity of β-Carotene

Mei Zi Sun  1 , Do-Yeong Kim  2 , Youjin Baek  1 , Hyeon Gyu Lee  1
Affiliations

The Effect of Multilayer Nanoemulsion on the In Vitro Digestion and Antioxidant Activity of β-Carotene

Mei Zi Sun et al. Antioxidants (Basel). .

Abstract

The objectives of this study were to design multilayer oil-in-water nanoemulsions using a layer-by-layer technique to enhance the stability of β-carotene and evaluate its effect on in vitro release and antioxidant activity. To prepare β-carotene-loaded multilayer nanoemulsions (NEs), a primary NE (PRI-NE) using Tween 20 was coated with chitosan (CS) for the secondary NE (SEC-CS), and with dextran sulfate (DS) and sodium alginate (SA) for the two types of tertiary NEs (TER-DS, TER-SA). The multilayer NEs ranged in particle size from 92 to 110 nm and exhibited high entrapment efficiency (92-99%). After incubation in a simulated gastrointestinal tract model, the release rate of free fatty acids decreased slightly after coating with CS, DS, and SA. The bioaccessibility of β-carotene was 7.02% for the PRI-NE, 7.96% for the SEC-CS, 10.88% for the TER-DS, and 10.25% for the TER-SA. The 2,2-diphenyl-1-picrylhydrazyl radical scavenging abilities increased by 1.2 times for the multilayer NEs compared to the PRI-NE. In addition, the cellular antioxidant abilities improved by 1.8 times for the TER-DS (87.24%) compared to the PRI-NE (48.36%). Therefore, multilayer nanoemulsions are potentially valuable techniques to improve the stability, in vitro digestion, and antioxidant activity of β-carotene.

Keywords: antioxidant; in vitro digestion; multilayer nanoemulsions; β-carotene.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Dependence of the particle size (a), PDI (a), DCR (b), and zeta potential (b) on chitosan concentration for primary nanoemulsions. Significant differences in the primary nanoemulsion’s particle size or zeta potential values depending on the concentration of CS were demonstrated with different capital letters (p < 0.05). Significant differences in the primary nanoemulsion’s PDI or DCR depending on the concentration of CS were demonstrated with different lowercase letters (p < 0.05). Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 2
Figure 2
Dependence of the particle size (a), PDI (a), DCR (b), and zeta potential (b) on dextran sulfate concentration for secondary nanoemulsions at a chitosan concentration of 0.25% (w/w). Significant differences in the primary nanoemulsion’s particle size or zeta potential values depending on the concentration of CS were demonstrated with different capital letters (p < 0.05). Significant differences in the primary nanoemulsion’s PDI or DCR depending on the concentration of CS were demonstrated with different lowercase letters (p < 0.05). Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 3
Figure 3
Dependence of the particle size (a), PDI (a), DCR (b), and zeta potential (b) on SA concentration for secondary nanoemulsions at a chitosan concentration of 0.25% (w/w). Significant differences in the primary nanoemulsion’s particle size or zeta potential values depending on the concentration of CS were demonstrated with different capital letters (p < 0.05). Significant differences in the primary nanoemulsion’s PDI or DCR depending on the concentration of CS were demonstrated with different lowercase letters (p < 0.05). Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 4
Figure 4
EE of PRI-NE, SEC-NE, TER-DS, and TER-SA. A Mean with letter is significantly different (p < 0.05). Key: PRI-NE = primary nanoemulsion; SEC-CS = secondary nanoemulsion coated with chitosan; TER-DS = tertiary nanoemulsion coated with dextran sulfate sodium salt; TER-SA = tertiary nanoemulsion coated with alginic acid sodium salt. Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 5
Figure 5
Cell viabilities of HEK293 cells treated with β-carotene-loaded multilayer nanoemulsions. Key: PRI-NE = primary nanoemulsion; SEC-CS = secondary nanoemulsion coated with chitosan; TER-DS = tertiary nanoemulsion coated with dextran sulfate sodium salt; TER-SA = tertiary nanoemulsion coated with alginic acid sodium salt. Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 6
Figure 6
Influence of simulated gastrointestinal conditions on the zeta potential of β-carotene-loaded multilayer nanoemulsions. Key: PRI-NE = primary nanoemulsion; SEC-CS = secondary nanoemulsion coated with chitosan; TER-DS = tertiary nanoemulsion coated with dextran sulfate sodium salt; TER-SA = tertiary nanoemulsion coated with alginic acid sodium salt. Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 7
Figure 7
Influence of simulated gastrointestinal conditions on the mean particle size of β-carotene-loaded multilayer nanoemulsions. Key: PRI-NE = primary nanoemulsion; SEC-CS = secondary nanoemulsion coated with chitosan; TER-DS = tertiary nanoemulsion coated with dextran sulfate sodium salt; TER-SA = tertiary nanoemulsion coated with alginic acid sodium salt. Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 8
Figure 8
Influence of the types of multilayer nanoemulsions on in vitro digestion under simulated small intestinal conditions. Key: PRI-NE = primary nanoemulsion; SEC-CS = secondary nanoemulsion coated with chitosan; TER-DS = tertiary nanoemulsion coated with dextran sulfate sodium salt; TER-SA = tertiary nanoemulsion coated with alginic acid sodium salt. Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 9
Figure 9
Influence of the types of multilayer nanoemulsions on the bioaccessibility (%) of β-carotene after in vitro digestion. A,B Means with different letters are significantly different (p < 0.05). Key: PRI-NE = primary nanoemulsion; SEC-CS = secondary nanoemulsion coated with chitosan; TER-DS = tertiary nanoemulsion coated with dextran sulfate sodium salt; TER-SA = tertiary nanoemulsion coated with alginic acid sodium salt. Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 10
Figure 10
The DPPH radical scavenging activities of β-carotene-loaded multilayer nanoemulsions. A–C Means with different letters are significantly different (p < 0.05). Key: PRI-NE = primary nanoemulsion; SEC-CS = secondary nanoemulsion coated with chitosan; TER-DS = tertiary nanoemulsion coated with dextran sulfate sodium salt; TER-SA = tertiary nanoemulsion coated with alginic acid sodium salt. Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
Figure 11
Figure 11
The CAA values of β-carotene-loaded multilayer nanoemulsions. A–C Means with different letters are significantly different (p < 0.05). Key: PRI-NE = primary nanoemulsion; SEC-CS = secondary nanoemulsion coated with chitosan; TER-DS = tertiary nanoemulsion coated with dextran sulfate sodium salt; TER-SA = tertiary nanoemulsion coated with alginic acid sodium salt. Measurement was performed in triplicate, and the results are expressed as the mean and standard deviation.
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