A Novel Sprouted Oat Fermented Beverage: Evaluation of Safety and Health Benefits for Celiac Individuals
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals, Reagents and Standards
2.2. Preparation of Sprouted Oat Flour
2.3. Bacterial Strain Activation and Inoculum Preparation
2.4. Manufacture of SOFB
2.5. Manufacture of Placebo Beverage for the Intervention Study
2.6. Analysis of Gluten Content
2.7. Evaluation of SOFB Protein Profile by Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE)
2.8. Determination of SOFB Immunotoxicity
2.9. Determination of SOFB Antioxidant and Antiinflammatory Activities
2.9.1. Cell Lines
2.9.2. In Vitro Gastrointestinal Digestion
2.9.3. Cell Treatments
2.9.4. Cell Viability
2.9.5. In Vitro Determination of Reactive Oxygen Species (ROS) Concentration
2.9.6. Anti-Inflammatory In Vitro Assay
2.10. In Vivo Evaluation of Safety and Health Benefits of SOFB in CD Individuals
2.10.1. Study Population and Design
2.10.2. Serological and Histological Analysis
2.10.3. Fecal Samples Collection and Metataxonomic Analysis
2.11. Statistical Analysis
3. Results
3.1. Evaluation of SOFB Gluten Content and Immunotoxicity
3.2. SOFB Did Not Protect Caco-2 Cells against t-BOOH-Induced Oxidative Stress
3.3. SOFB Attenuates Inflammatory Response in LPS-Induced RAW 264.7 Macrophages
3.4. In Vivo Evaluation of SOFB Safety and Health Benefits in CD Subjects
3.4.1. Patient Baseline Characteristics
3.4.2. SOFB Safety Assessment in CD Subjects
3.4.3. SOFB Effects on Body Weight and Serological Markers of Nutritional Status in CD Subjects
3.4.4. Metataxonomic Analysis of Intestinal Microbiota in CD Subjects
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Baseline Characteristics | Control Group (n = 5) | SOFB Group (n = 5) | p |
---|---|---|---|
Gender (male/female) | 2/3 | 2/3 | |
Age (year) | 45.20 ± 12.32 | 31.80 ± 12.52 | 0.126 |
Duration GF diet (year) | 8.30 ± 9.36 | 8.80 ± 6.69 | 0.530 |
Weight (Kg) | 72.20 ± 14.81 | 62.80 ± 5.80 | 0.168 |
Height (m) | 1.68 ± 0.08 | 1.68 ± 0.08 | 0.727 |
Body mass index (Kg/m2) | 25.54 ± 4.00 | 21.50 ± 1.99 | 0.078 |
IgA Levels | Control Group | SOFB Group | p |
---|---|---|---|
Total serum IgA (mg/dL) | |||
0 months | 200.00 ± 67.99 (146.0–288.0) a | 197.20 ± 60.26 (104.0–271.0) a | 1.000 |
2 months | 188.40 ± 70.18 (126.0–286.0) a | 192.34 ± 61.95 (99.7–270.0) a | 1.000 |
4 months | 196.00 ± 67.66 (143.0–288.0) a | 200.80 ± 66.38 (104.0–290.0) a | 0.841 |
6 months | 202.20 ± 79.90 (1.26–311.0) a | 201.80 ± 67.15 (102.0–290.0) a | 1.000 |
IgA-tTG (U/mL) | |||
0 months | 1.76 ± 0.62 (0.7–2.4) a | 2.72 ± 3.29 (0.5–8.5) a | 0.841 |
2 months | 1.38 ± 0.62 (0.5–2.0) a | 2.64 ± 3.03 (0.9–8.0) a | 1.000 |
4 months | 1.83 ± 0.54 (1.0–2.4) a | 2.39 ± 2.41 (1.0–6.7) a | 0.548 |
6 months | 1.22 ± 0.56 (0.5–1.8) a | 2.55 ± 2.74 (0.2–7.2) a | 0.421 |
IEL Levels | Months | Control Group | SOFB Group | p | |
---|---|---|---|---|---|
Second duodenal portion | Total IEL | 0 | 10.80 ± 2.83 (6.80–12.90) a | 19.90 ± 9.84 (5.20–25.80) a | 0.343 |
6 | 8.70 ± 4.04 (3.90–12.10) a | 8.24 ± 6.37 (2.50–0.40) a | 0.841 | ||
TCR γδ+ IEL | 0 | 28.28 ± 12.41 (19.40–46.50) a | 24.32 ± 9.55 (17.00–40.90) a | 0.413 | |
6 | 22.98 ± 6.77 (16.30–31.60) a | 29.16 ± 12.20 (16.50–46.70) a | 0.548 | ||
CD3−CD103+ IEL | 0 | 6.10 ± 5.74 (0.30–13.20) a | 3.12 ± 3.41 (0.40–8.40) a | 0.730 | |
6 | 7.44 ± 9.09 (1.20–23.50) a | 3.20 ± 4.15 (1.10–10.60) a | 0.222 | ||
Duodenal bulb | Total IEL | 0 | 8.30 ± 4.51 (4.20–14.40) a | 11.57 ± 7.75 (5.9–20.40) a | 0.533 |
6 | 5.83 ± 1.28 (4.40–7.40) a | 3.97 ± 2.35 (1.40–6.00) a | 0.800 | ||
TCR γδ+ IEL | 0 | 19.00 ± 15.67 (6.90–40.90) a | 24.46 ± 12.12 (14.90–38.10) a | 0.630 | |
6 | 20.80 ± 17.81 (8.20–47.20) a | 24.70 ± 10.07 (14.30–34.40) a | 0.533 | ||
CD3−CD103+ IEL | 0 | 3.23 ± 1.61 (1.80–4.90) a | 3.30 ± 4.33 (0.70–8.30) a | 0.629 | |
6 | 1.98 ± 0.86 (1.20–3.20) a | 1.96 ± 0.83 (1.30–2.90) a | 0.857 |
IgA Levels | Control Group | SOFB Group | p |
---|---|---|---|
Body weight (Kg) | |||
0 months | 72.2 ± 14.8 (54.0–95.0) a | 62.8 ± 5.8 (54.0–68.0) a | 0.151 |
2 months | 73.5 ± 16.8 (54.5–100.0) a | 62.7 ± 6.1 (52.5–68.0) a | 0.151 |
4 months | 73.5 ± 16.4 (54.0–99.0) a | 63.1 ± 6.4 (52.5–69.0) a | 0.095 |
6 months | 73.5 ± 16.7 (52.5–99.0) a | 62.8 ± 7.2 (51.0–70.0) a | 0.151 |
Glucose (μg/dL) | |||
0 months | 89.4 ± 7.4 (79.0–97.0) a | 80.4 ± 6.9 (72.0–87.0) a | 0.151 |
2 months | 83.8 ± 6.5 (73.0–90.0) a | 76.6 ± 10.7 (64.0–92.0) a | 0.222 |
4 months | 88.2 ± 12.1 (75.0–106.0) a | 83.2 ± 1.7 (82.0–86.0) a | 0.690 |
6 months | 84.2 ± 12.0 (71.0–95.0) a | 80.8 ± 5.9 (71.0–85.0) a | 0.421 |
Iron (mg/dL) | |||
0 months | 98.6 ± 22.1 (79.0–132.0) a | 100.0 ± 26.5 (73.0–127.0) a | 0.841 |
2 months | 86.2 ± 16.6 (64.0–108.0) a | 104.4 ± 33.8 (63.0–145.0) a | 0.420 |
4 months | 88.0 ± 27.2 (53.0–119.0) a | 69.6 ± 43.8 (38.0–145.0) a | 0.222 |
6 months | 101.6 ± 8.0 (95.0–114.0) a | 97.4 ± 61.7 (44.0–189.0) a | 0.690 |
Ferritin (ng/mL) | |||
0 months | 62.9 ± 49.2 (20.9–143.5) a | 80.9 ± 66.5 (24.1–167.9) a | 0.917 |
2 months | 73.6 ± 59.0 (17.3–164.9) a | 72.0 ± 60.1 (18.4–154.3) a | 0.917 |
4 months | 72.1 ± 81.2 (19.1–214.0) a | 76.5 ± 69.4 (15.6–177.7) a | 0.917 |
6 months | 80.5 ± 96.7 (13.1–247.3) a | 76.3 ± 69.2 (17.3–177.7) a | 0.754 |
Vitamin B12 (pg/mL) | |||
0 months | 416.2 ± 143.8 (261.0–610.0) a | 407.8 ± 120.1 (261.0–589.0) a | 0.690 |
2 months | 428.2 ± 164.2 (246.0–644.0) a | 398.0 ± 136.8 (217.0–589.0) a | 1.000 |
4 months | 449.0 ± 202.7 (167.0–610.0) a | 393.8 ± 123.3 (218.0–544.0) a | 0.548 |
6 months | 387.6 ± 121.4 (261.0–527.0) a | 417.2 ± 186.7 (274.0–728.0) a | 0.841 |
Folic acid (ng/mL) | |||
0 months | 7.5 ± 3.7 (3.7–13.2) a | 5.1 ± 2.6 (2.8–9.4) a | 0.310 |
2 months | 9.8 ± 4.2 (6.1–15.9) a | 5.7 ± 3.0 (2.1–9.0) a | 0.222 |
4 months | 6.6 ± 1.9 (4.5–8.8) a | 4.9 ± 2.4 (2.1–8.7) a | 0.220 |
6 months | 8.0 ± 2.2 (5.4–10.7) a | 5.1 ± 2.1 (3.1–8.7) a | 0.032 |
Hemoglobin (g/dL) | |||
0 months | 14.4 ± 1.3 (12.5–16.2) a | 14.1 ± 0.8 (13.3–15.2) a | 0.690 |
2 months | 14.4 ± 1.5 (12.8–16.7) a | 12.0 ± 4.3 (4.4–14.3) a | 0.310 |
4 months | 14.3 ± 1.3 (12.8–16.0) a | 12.1 ± 4.1 (4.9–14.3) a | 0.548 |
6 months | 14.2 ± 1.9 (11.9–16.4) a | 13.9 ± 0.4 (13.3–14.3) a | 0.841 |
Triglycerides (mg/mL) | |||
0 months | 133.8 ± 43.6 (64.0–174.0) a | 53.6 ± 19.3 (34.0–82.0) a | 0.016 |
2 months | 110.8 ± 47.8 (68.0–190.0) a | 69.6 ± 46.0 (33.0–150.0) a | 0.095 |
4 months | 94.8 ± 26.9 (64.0–130.0) a | 81.0 ± 48.0 (30.0–150.0) a | 0.069 |
6 months | 116.4 ± 59.7 (51.0–207.0) a | 51.6 ± 20.4 (41.0–88.0) a | 0.016 |
Total cholesterol (mg/dL) | |||
0 months | 198.0 ± 41.2 (147.0–262.0) a | 147.2 ± 22.4 (119.0–170.0) a | 0.060 |
2 months | 188.2 ± 25.9 (158.0–215.0) a | 132.0 ± 28.4 (94.0–163.0) a | 0.016 |
4 months | 189.0 ± 42.3 (147.0–260.0) a | 137.4 ± 18.32 (115.0–158.0) a | 0.032 |
6 months | 199.0 ± 30.2 (163.0–235.0) a | 146.6 ± 21.4 (124.0–175.0) a | 0.016 |
Phylum/Genus | Control Group | SOFB Group | p | ||
---|---|---|---|---|---|
n (%) | Median (IQR) | n (%) # | Median (IQR) | ||
Firmicutes | 20 (100%) | 88.78 (80.85–92.2) | 20 (100%) | 91.46 (89.03–93.52) | 0.081 |
Blautia | 20 (100%) | 9.79 (7–12.58) | 20 (100%) | 10.07 (6.32–14.58) | 0.95 |
Subdoligranulum | 20 (100%) | 2.72 (1.42–6.05) | 20 (100%) | 6.99 (4.72–8.93) | 0.002 |
Faecalibacterium | 16 (80%) | 3.83 (0.64–7.9) | 20 (100%) | 3.85 (1.61–8.07) | 0.6 |
Agathobacter | 20 (100%) | 2.04 (1.58–5.25) | 20 (100%) | 1.96 (1.18–7.05) | 0.82 |
Dorea | 20 (100%) | 3.19 (2.56–5.56) | 20 (100%) | 3.87 (1.6–5.11) | 0.56 |
Anaerostipes | 20 (100%) | 3.41 (1.19–8.27) | 20 (100%) | 1.34 (0.6–3.13) | 0.02 |
Streptococcus | 20 (100%) | 1.9 (1.23–4.22) | 20 (100%) | 1.11 (0.54–3.44) | 0.15 |
Erysipelotrichaceae | 20 (100%) | 4.02 (1.75–4.83) | 19 (95%) | 2.61 (0.66–4.04) | 0.12 |
Ruminococcus | 6 (30%) | <0.01 (<0.01–0.01) | 17 (85%) | 5.28 (0.16–7.17) | <0.001 |
Lactobacillus | 15 (75%) | 0.04 (<0.01–0.13) | 17 (85%) | 0.25 (0.05–3.11) | 0.028 |
Clostridium | 15 (75%) | 0.4 (0.01–1.19) | 20 (100%) | 0.38 (0.18–2.08) | 0.25 |
Romboutsia | 17 (85%) | 0.78 (0.38–1.48) | 20 (100%) | 1.02 (0.43–2.62) | 0.39 |
Holdemanella | 4 (20%) | <0.01 (<0.01–<0.01) | 8 (40%) | <0.01 (<0.01–1.56) | 0.43 |
Monoglobus | 16 (80%) | 1.29 (0.1–1.89) | 19 (95%) | 0.95 (0.3–2.32) | 0.64 |
Coprococcus | 17 (85%) | 1.35 (0.98–1.8) | 20 (100%) | 1.8 (1.17–2.1) | 0.13 |
Catenibacterium | 5 (25%) | <0.01 (<0.01–<0.01) | 4 (20%) | <0.01 (<0.01–<0.01) | 0.81 |
Actinobacteria | 20 (100%) | 5.69 (4.41–6.87) | 20 (100%) | 5.43 (3.58–6.51) | 0.24 |
Bifidobacterium | 20 (100%) | 1.96 (0.79–4.09) | 19 (95%) | 1.72 (0.52–3.16) | 0.58 |
Bacterioidota | 20 (100%) | 3.01 (0.86–11.09) | 19 (95%) | 2.17 (0.58–4.13 | 0.3 |
Bacteroides | 18 (90%) | 2.5 (0.61–7.67) | 18 (90%) | 1.24 (0.25–2.58) | 0.16 |
Proteobacteria | 18 (90%) | 0.17 (0.05–0.67) | 20 (100%) | 0.25 (0.14–0.64) | 36 |
Fusobacteriota | 10 (50%) | 0.01 (<0.01–0.11) | 3 (15%) | <0.01 (<0.01–<0.01) | 0.011 |
Minor phyla | 19 (95%) | 0.07 (0.02–0.21) | 16 (80%) | 0.06 (0.01–0.6) | 0.86 |
Minor genera | 20 (100%) | 15.44 (14.57–21.34) | 20 (100%) | 17.55 (13.63–21.97) | 0.97 |
Unclassified genera | 20 (100%) | 17.55 (13.63–21.97) | 20 (100%) | 13.3 (10.94–14.75) | 0.001 |
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Aparicio-García, N.; Martínez-Villaluenga, C.; Frias, J.; Crespo Perez, L.; Fernández, C.F.; Alba, C.; Rodríguez, J.M.; Peñas, E. A Novel Sprouted Oat Fermented Beverage: Evaluation of Safety and Health Benefits for Celiac Individuals. Nutrients 2021, 13, 2522. https://doi.org/10.3390/nu13082522
Aparicio-García N, Martínez-Villaluenga C, Frias J, Crespo Perez L, Fernández CF, Alba C, Rodríguez JM, Peñas E. A Novel Sprouted Oat Fermented Beverage: Evaluation of Safety and Health Benefits for Celiac Individuals. Nutrients. 2021; 13(8):2522. https://doi.org/10.3390/nu13082522
Chicago/Turabian StyleAparicio-García, Natalia, Cristina Martínez-Villaluenga, Juana Frias, Laura Crespo Perez, Cristina Fernández Fernández, Claudio Alba, Juan Miguel Rodríguez, and Elena Peñas. 2021. "A Novel Sprouted Oat Fermented Beverage: Evaluation of Safety and Health Benefits for Celiac Individuals" Nutrients 13, no. 8: 2522. https://doi.org/10.3390/nu13082522