New Perspectives on the Sustainable Employment of Chestnut Shells as Active Ingredient against Oral Mucositis: A First Screening
Abstract
:1. Introduction
2. Results and Discussion
2.1. Extraction Yield
2.2. Extraction Yield
2.3. Phenolic Profile Identification and Quantification by HPLC-PDA
2.4. ROS Scavenging Capacity
2.5. Antimicrobial Activity
2.6. Cells Viability Assays
3. Materials and Methods
3.1. Chemicals
3.2. Sample
3.3. Subcritical Water Extraction (SWE) of C. sativa Shells
3.4. Determination of Total Phenolic Content
3.5. Antioxidant and Antiradical Activities
3.5.1. Ferric Reducing Antioxidant Power (FRAP) Assay
3.5.2. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) Free Radical Scavenging Assay
3.5.3. 2,2´-Azino-bis-3-ethylbenzothiazoline-6-sulfonic Acid (ABTS) Radical Scavenging Activity Assay
3.6. HPLC-PDA Analysis
3.7. Reactive Oxygen Species and Reactive Nitrogen Species Scavenging Capacity
3.7.1. Superoxide Radical Scavenging Assay
3.7.2. Hypochlorous Acid Scavenging Assay
3.7.3. Peroxyl Radical Scavenging Assay
3.7.4. Peroxynitrite Scavenging Assay
3.8. Antimicrobial Activity
3.8.1. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)
3.8.2. Growth Rate (GR) Studies
3.8.3. Minimum Bactericidal Concentration (MBC) Determination
3.9. Cell Viability Assay
3.10. Statistical Analysis
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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SWE Extracts | Extraction Yield (%) | TPC (mg GAE/g DW) | DPPH IC50 (µg/mL) | ABTS IC50 (µg/mL) | FRAP (µmol FSE/g) |
---|---|---|---|---|---|
110 °C | 20.88 ± 0.78 | 239.53 ± 23.17 a | 426.88 ± 13.42 | 148.68 ± 13.20 a | 4240.38 ± 10.04 a |
120 °C | 21.00 ± 1.49 | 201.75 ± 13.02 b | 489.49 ± 5.96 | 228.32 ± 16.48 b | 4092.98 ± 92.82 b |
140 °C | 20.83 ± 0.62 | 162.52 ± 7.14 c | 460.21 ± 14.81 | 232.97 ± 4.23 b | 3398.98 ± 57.23 c |
160 °C | 20.97 ± 1.74 | 122.31 ± 5.89 d | 496.15 ± 8.75 | 220.40 ± 7.83 b | 2728.06 ± 35.04 d |
180 °C | 20.29 ± 0.93 | 126.19 ± 6.33 d | 583.47 ± 14.68 | 256.59 ± 5.55 b | 2464.32 ± 68.44 e |
Phenolic Compound | 110 °C | 120 °C | 140 °C | 160 °C | 180 °C |
---|---|---|---|---|---|
mg/g DW | |||||
Alkaloids | |||||
Caffeine | 1.18 ± 0.06 | 1.14 ± 0.06 | 1.01 ± 0.05 | 0.33 ± 0.02 | 0.44 ± 0.02 |
Chalconoids | |||||
Phloridzin | 0.83 ± 0.04 | 0.50 ± 0.03 | ND | ND | ND |
Flavanols | |||||
Catechin | 4.10 ± 0.20 | 3.65 ± 0.18 | 1.60 ± 0.08 | 0.74 ± 0.04 | 0.69 ± 0.03 |
Epicatechin | 1.68 ± 0.08 | 1.14 ± 0.06 | 0.75 ± 0.04 | <LOQ | <LOQ |
Flavonols | |||||
Rutin | 1.04 ± 0.05 | 0.64 ± 0.03 | 0.69 ± 0.03 | 0.103 ± 0.005 | 0.136 ± 0.007 |
Flavonones | |||||
Naringin | <LOQ | <LOQ | ND | ND | ND |
Phenolic acids | |||||
3,5-di-caffeolquinic acid | <LOQ | <LOQ | <LOQ | ND | ND |
4-O-caffeyolquinic acid | 0.49 ± 0.02 | 0.46 ± 0.02 | 0.40 ± 0.02 | 0.34 ± 0.02 | 0.21 ± 0.01 |
4,5-DI-O-caffeoylquinic acid | 0.126 ± 0.006 | 0.22 ± 0.01 | 0.34 ± 0.02 | 0.026 ± 0.001 | 0.28 ± 0.01 |
Caftaric acid | 0.44 ± 0.02 | 0.26 ± 0.01 | 0.53 ± 0.03 | 0.16 ± 0.01 | 0.58 ± 0.03 |
Caffeic acid | 0.17 ± 0.01 | 0.16 ± 0.01 | 0.12 ± 0.01 | 0.069 ± 0.003 | 0.12 ± 0.01 |
Clorogenic acid | 0.43 ± 0.02 | 0.27 ± 0.01 | 0.18 ± 0.01 | 0.081 ± 0.004 | 0.13 ± 0.01 |
p-Coumaric acid | <LOD | <LOD | <LOD | <LOD | <LOD |
Ellagic acid | 0.136 ± 0.007 | 0.157 ± 0.008 | 0.181 ± 0.009 | 0.064 ± 0.003 | 0.108 ± 0.005 |
Gallic acid | 5.99 ± 0.30 | 5.08 ± 0.25 | 4.64 ± 0.23 | 3.48 ± 0.17 | 2.40 ± 0.12 |
Ferulic acid | <LOQ | <LOQ | <LOQ | 0.011 ± 0.001 | 0.056 ± 0.003 |
Neochlorogenic acid | 0.71 ± 0.04 | 0.59 ± 0.03 | 0.49 ± 0.02 | 0.21 ± 0.01 | 0.28 ± 0.01 |
Protocatechuic acid | 1.68 ± 0.08 | 1.37 ± 0.07 | 1.15 ± 0.06 | 1.31 ± 0.07 | 1.59 ± 0.08 |
Sinapic acid | ND | ND | ND | <LOQ | <LOQ |
Syringic acid | ND | ND | ND | 0.019 ± 0.001 | 0.063 ± 0.003 |
Vanillic acid | 1.02 ± 0.05 | 0.91 ± 0.05 | 0.55 ± 0.03 | 0.084 ± 0.004 | 0.12 ± 0.01 |
Quercetin-3-O-galactoside | 0.115 ± 0.006 | 0.099 ± 0.005 | 0.136 ± 0.007 | 0.012 ± 0.001 | 0.120 ± 0.006 |
Quercetin-3-O-glucopyranoside | 0.057 ± 0.003 | 0.069 ± 0.003 | 0.081 ± 0.004 | 0.043 ± 0.002 | 0.036 ± 0.002 |
Stilbenoids | |||||
Resveratrol | <LOD | <LOD | <LOD | ND | ND |
Trans-polydatin | <LOQ | <LOD | <LOD | <LOD | <LOD |
Total | 20.20 | 16.71 | 12.85 | 7.07 | 7.37 |
Reactive Oxygen Species | Reactive Nitrogen Species | ||||
---|---|---|---|---|---|
SWE Extracts | O2•− | HOCl | ROO• | ONOO- in Presence of HCO3− | ONOO- in Absence of HCO3− |
IC50 (µg/mL) | IC50 (µg/mL) | (µmol TE/mg DW) | IC50 (µg/mL) | ||
110 °C | 44.36 ± 0.34 cd | 4.47 ± 0.29 b | 0.73 ± 0.024 b | 2.52 ± 0.19 c | 2.53 ± 0.22 bc |
120 °C | 46.31 ± 1.37 d | 6.49 ± 0.18 b | 0.54 ± 0.016 cd | 1.41 ± 0.11 b | 3.32 ± 0.19 c |
140 °C | 31.14 ± 2.05 b | 8.27 ± 0.77 b | 0.61 ± 0.037 bc | 2.77 ± 0.29 c | 2.09 ± 0.21 b |
160 °C | 35.34 ± 3.14 bc | 22.85 ± 1.67 c | 0.50 ± 0.032 cd | 2.79 ±0.26 c | 1.89 ± 0.09 b |
180 °C | 73.18 ±1.86 e | 20.74 ± 1.27 c | 0.49 ± 0.009 d | 2.93 ± 0.03 c | 5.94 ± 0.30 d |
Positive control | |||||
Gallic acid | 23.82 ± 0.82 a | 1.68 ± 0.16 a | 4.23 ± 0.25 a | 0.30 ± 0.03 a | 0.05 ± 0.00 a |
SWE Extracts | Gram-Negative | Gram-Positive | |||||
---|---|---|---|---|---|---|---|
E. coli ATCC 25922 | E. coli ATCC 8739 | E. coli CTX M2 * | S. aureus ATCC 25913 | S. aureus MRSA * | E. faecalis | ||
110 °C | MIC | 8 | 4 | 4 | - | 4 | 64 |
MBC | 64 | - | - | - | 32 | - | |
120 °C | MIC | 8 | 4 | 8 | 64 | 8 | 2 |
MBC | 64 | - | - | - | 16 | - | |
140 °C | MIC | 4 | 16 | 4 | 4 | 32 | 64 |
MBC | - | - | - | - | 64 | - | |
160 °C | MIC | 4 | 4 | 4 | 4 | 4 | 8 |
MBC | 32 | - | - | 64 | 32 | 64 | |
180 °C | MIC | 8 | 8 | - | 4 | 4 | 16 |
MBC | 16 | 4 | - | 64 | 16 | 64 |
Cell Lines IC50 (µg/mL) | ||
---|---|---|
SWE Extracts | HSC3 | TR146 |
110 °C | 1325.03 | 468.15 |
120 °C | 1730.39 | 496.45 |
140 °C | 1862.17 | 663.83 |
160 °C | 1923.94 | 553.79 |
180 °C | 1707.33 | 572.18 |
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Ferreira, A.S.; Silva, A.M.; Pinto, D.; Moreira, M.M.; Ferraz, R.; Švarc-Gajić, J.; Costa, P.C.; Delerue-Matos, C.; Rodrigues, F. New Perspectives on the Sustainable Employment of Chestnut Shells as Active Ingredient against Oral Mucositis: A First Screening. Int. J. Mol. Sci. 2022, 23, 14956. https://doi.org/10.3390/ijms232314956
Ferreira AS, Silva AM, Pinto D, Moreira MM, Ferraz R, Švarc-Gajić J, Costa PC, Delerue-Matos C, Rodrigues F. New Perspectives on the Sustainable Employment of Chestnut Shells as Active Ingredient against Oral Mucositis: A First Screening. International Journal of Molecular Sciences. 2022; 23(23):14956. https://doi.org/10.3390/ijms232314956
Chicago/Turabian StyleFerreira, Ana Sofia, Ana Margarida Silva, Diana Pinto, Manuela M. Moreira, Ricardo Ferraz, Jaroslava Švarc-Gajić, Paulo C. Costa, Cristina Delerue-Matos, and Francisca Rodrigues. 2022. "New Perspectives on the Sustainable Employment of Chestnut Shells as Active Ingredient against Oral Mucositis: A First Screening" International Journal of Molecular Sciences 23, no. 23: 14956. https://doi.org/10.3390/ijms232314956