journal article·2025-10-28Open access

The Effect of In Vitro Gastrointestinal Digestion on the Biological Activity of a Sea Cucumber (Holothuria forskali) Hydrolysate Encapsulated in Chitosan Nanoparticles and Rapeseed Lecithin Liposomes

Ailén Alemán, María del Carmen Gómez-Guillén, María Pilar Montero, María Elvira López-Caballero

Applied Sciences· Volume 15· Issue 21

Abstract

A growing scientific interest in bioactive compounds from sea cucumbers is contributing to a broader recognition even in regions where their consumption is not common. This study evaluated the biological potential of a Holothuria forskali extract obtained through different extraction methods, including water extraction, ethanol–water extraction, and enzymatic hydrolysis. The hydrolysate (H), rich in low-molecular-weight peptides, yielded the highest antioxidant (30.6 ± 0.6 mg VitC Eq/g sample for ABTS and 10.7 ± 0.1 mg GAEs/g sample for Folin-reactive substances) and ACE-inhibitory (82.6%) activities. Based on these results, the hydrolysate was selected for encapsulation in two nanostructured delivery systems for comparative purposes: chitosan nanoparticles (NPs) and rapeseed lecithin liposomes (LPs). Both nanostructures were characterized in terms of size, ζ-potential, and polydispersity and subjected to simulated in vitro gastrointestinal digestion (GIDv) to assess their stability and mucoadhesive properties. After digestion, antioxidant activity increased in both systems, particularly in liposomes. Although encapsulation initially reduced ACE-inhibitory activity, gastrointestinal digestion restored or enhanced it, especially in liposomal formulations (≈37% inhibition). The mucoadhesive potential of the nanostructures after DGIv, focusing on their interactions with mucin, was assessed. Liposomal digests significantly increased viscosity in the presence of mucin, while chitosan nanoparticles decreased it, suggesting the formation of soluble complexes with reduced hydrodynamic volume. Electrostatic and hydrogen bonding interactions between chitosan and mucin were particularly evident in the NPH formulation. The rheological synergism parameter (Δη) revealed more negative values for NPs and NPHs, indicating stronger mucoadhesive interactions compared to controls and suggesting their suitability for mucosal delivery. These findings support the use of H. forskali hydrolysates as a source of functional bioactive compounds and highlight the potential of chitosan-based nanocarriers for enhancing their stability, bioaccessibility, and mucoadhesive properties in functional food or nutraceutical applications.
Source: crossref · Quality: baseline

Research context

OpenAlex topics: Echinoderm biology and ecology · Seaweed-derived Bioactive Compounds · Protein Hydrolysis and Bioactive Peptides

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Institute of Food Science, Technology and Nutrition (ICTAN-CSIC), C/José Antonio Novais 6, 28040 Madrid, Spain

References (83)

Show source-provided references
1. (2025, February 04). UN. Available online: http://www.un.org>global-issues>ageing.
2. High-Value Components and Bioactives from Sea Cucumbers for Functional Foods—A Review10.3390/md9101761 ↗
3. Ismail, S.-E., Hussein, N.A., Rashad, M.M., El-Sikaily, A.M., Hassanin, A.E.-L.A., and Fakharany, E.M.E. (2025). Sea cucumber sulphated polysaccharides extract potentiates the anticancer effect of 5- fluorouracil on hepatocellular carcinoma cells. Sci. Rep., 15.10.1038/s41598-025-06496-7 ↗
4. Hossain, A., Dave, D., and Shahidi, F. (2020). Northern Sea Cucumber (Cucumaria frondosa): A Potential Candidate for Functional Food, Nutraceutical, and Pharmaceutical Sector. Mar. Drugs, 18.10.3390/md18050274 ↗
5. The neuropeptidomes of the sea cucumbers Stichopus cf. horrens and Holothuria scabra10.1038/s41598-025-85696-7 ↗
6. Shahbaz, M., Akram, A., Raja, N.I., Mukhtar, T., Mehak, A., Fatima, N., Ajmal, M., Ali, K., Mustafa, N., and Abasi, F. (2023). Antifungal activity of green synthesized selenium nanoparticles and their effect on physiological, biochemical, and antioxidant defense system of mango under mango malformation disease. PLoS ONE, 18.10.1371/journal.pone.0274679 ↗
7. Food emulsions and foams: Stabilization by particles10.1016/j.cocis.2009.11.001 ↗
8. Lima, K.O., da Rocha, M., Alemán, A., López-Caballero, M.E., Tovar, C.A., Gómez-Guillén, M.C., Montero, P., and Prentice, C. (2021). Yogurt fortification by the addition of microencapsulated strippedweakfish (Cynoscion guatucupa) protein hydrolysate. Antioxidants, 10.10.3390/antiox10101567 ↗
9. Carboxymethyl cellulose films containing nanoliposomes loaded with an angiotensin-converting enzyme inhibitory collagen hydrolysate10.1016/j.foodhyd.2019.04.009 ↗
10. Alemán, A., Pérez-García, S., Fernández de Palencia, P., Montero, M.P., and Gómez-Guillén, M.D.C. (2021). Physicochemical, antioxidant, and anti-inflammatory properties of rapeseed lecithin liposomes loading a chia (Salvia hispanica L.) seed extract. Antioxidants, 10.10.3390/antiox10050693 ↗
11. Comparative assessment of plant lecithins as the technologically functional components for creating encapsulated biologically active substances
12. Mecheta, A., Hanachi, A., Jeandel, C., Arab-Tehrany, E., Bianchi, A., Velot, E., Mezali, K., and Linder, M. (2020). Physicochemical Properties and Liposomal Formulations of Hydrolysate Fractions of Four Sea Cucumbers (Holothuroidea: Echinodermata) from the Northwestern Algerian Coast. Molecules, 25.10.3390/molecules25132972 ↗
13. Sea cucumber saponin liposomes ameliorate obesity-induced inflammation and insulin resistance in high-fat-diet-fed mice10.1039/C7FO01599B ↗
14. Transport and uptake effects of marine complex lipid liposomes in small intestinal epithelial cell models10.1039/C6FO00066E ↗
15. Characteristics of chitosan nanoparticles extracted from sea cucumber (Holothuria scabra) as source materials for glucosamine
16. Citkowska, A., Szekalska, M., and Winnicka, K. (2019). Possibilities of fucoidan utilization in the development of pharmaceutical dosage forms. Mar. Drugs, 17.10.3390/md17080458 ↗
17. Characterization and Absorption Kinetics of a Novel Multifunctional Nanoliposome Stabilized by Sea Cucumber Saponins Instead of Cholesterol10.1021/acs.jafc.9b06460 ↗
18. Preparation and properties of fucoxanthin-loaded liposomes stabilized by sea cucumber derived cholesterol sulfate instead of cholesterol10.1016/j.jbiosc.2022.11.004 ↗
19. Preparation and characterization of astaxanthin-loaded liposomes stabilized by sea cucumber sulfated sterols instead of cholesterol10.5650/jos.ess21233 ↗
20. Nanoliposomal Cercodemasoide A and Its Improved Activities Against NTERA-2 Cancer Stem Cells
21. Exploring in vitro effect of silver nanoparticles and Holothuria parva extracts on kinetics and stability of α-amylase10.1002/bab.2407 ↗
22. A rapid method of total lipid extraction and purification10.1139/y59-099 ↗
23. Quantification of phenolic contents and antioxidant capacity of Atlantic sea cucumber, Cucumaria frondosa10.1016/j.foodchem.2007.01.016 ↗
24. Antioxidant activity of several marine skin gelatins10.1016/j.lwt.2010.09.003 ↗
25. A Novel Functional Wrapping Design by Complexation of ε-Polylysine with Liposomes Entrapping Bioactive Peptides10.1007/s11947-016-1703-4 ↗
26. Exploring the potential of common iceplant, seaside arrowgrass and sea fennel as edible halophytic plants10.1016/j.foodres.2020.109613 ↗
27. Comparative study between film and coating packaging based on shrimp concentrate obtained from marine industrial waste for fish sausage preservation10.1016/j.foodcont.2016.06.007 ↗
28. A standardised static in vitro digestion method suitable for food—An international consensus10.1039/C3FO60702J ↗
29. Evisceration of Holothuria poli by mechanical, chemical and hypoxia stress methods and its bioremediation potentials for the pisciculture wastewater10.1111/are.15838 ↗
30. Compositional characteristics and antioxidant properties of fresh and processed sea cucumber (Cucumaria frondosa)10.1021/jf063085h ↗
31. The changes in chemical composition of Holothuria tubulosa (Gmelin, 1788) with ambient-drying and oven-drying methods10.1002/fsn3.703 ↗
32. Carletti, A., Cardoso, C., Lobo-Arteaga, J., Sales, S., Juliao, D., Ferreira, I., Chainho, P., Dionísio, M.A., Gaudêncio, M.J., and Afonso, C. (2022). Antioxidant and Anti-inflammatory Extracts From Sea Cucumbers and Tunicates Induce a Pro-osteogenic Effect in Zebrafish Larvae. Front. Nutr., 9.10.3389/fnut.2022.888360 ↗
33. Antioxidant capacity of dry sea cucumber Holothuria edulis, Pearsonothuria graeffei, and Stichopus herrmanni from Boalemo waters, Gorontalo, Indonesia
34. In Vitro Antioxidant and Antiproliferative Activities of Three Malaysian Sea Cucumber Species
35. Antioxidant and cytotoxic properties of two sea cucumbers, Holothuria edulis Lesson and Stichopus horrens Selenka10.1556/ABiol.64.2013.1.2 ↗
36. Antioxidant properties of green tea extract incorporated to fish gelatin films after simulated gastrointestinal enzymatic digestion10.1016/j.lwt.2013.03.020 ↗
37. Changes in phenolic composition and antioxidant activities of the edible halophyte Crithmum maritimum L. with physiological stage and extraction method10.1007/s11738-012-0943-9 ↗
38. First report of the nutritional profile and antioxidant potential of Holothuria arguinensis, a new resource for aquaculture in Europe10.1080/14786419.2015.1107555 ↗
39. Antioxidant potential and physicochemical properties of protein hydrolysates from body parts of North Atlantic sea cucumber (Cucumaria frondosa)10.1186/s43014-020-00049-3 ↗
40. Angiotensin-I Converting Enzyme (ACE) Inhibitory and Anti-Oxidant Activities of Sea Cucumber (Actinopyga lecanora) Hydrolysates10.3390/ijms161226140 ↗
41. Antioxidant and antibacterial activities in 21 species of Indonesian sea cucumbers10.1007/s13197-021-05007-6 ↗
42. Antibacterial activities and bioactive compounds of the ethyl acetate extract of the sea cucumber Holothuria forskali from Tunisian coasts10.1017/S0025315420000016 ↗
43. Antimicrobial substances of potential biomedical importance from holothurian species
44. Antifungal activity in triterpene glycosides from the sea cucumber Actinopyga lecanora10.1016/j.bmcl.2006.12.052 ↗
45. Characterization of phenoloxidase from the sea cucumber Apostichopus japonicas10.1016/j.imbio.2014.02.006 ↗
46. Actinopyga lecanora Hydrolysates as Natural Antibacterial Agents10.3390/ijms131216796 ↗
47. Production of bioactive peptides from sea cucumber and its potential health benefits: A comprehensive review10.1021/acs.jafc.2c02696 ↗
48. Bioactive compounds, antioxidant potential, and hepatoprotective activity of sea cucumber (Holothuria atra) against thioacetamide intoxication in rats10.1016/j.nut.2012.06.004 ↗
49. Study on the free radical scavenging activity of sea cucumber (Paracaudina chinens var.) gelatin hydrolysate10.1007/s11802-007-0255-7 ↗
50. Antioxidant activity and sensory quality of asparagus affected by microwave-circulated water combination and conventional cooking methods
51. Antihypertensive effect and purification of an ACE inhibitory peptide from sea cucumber gelatin hydrolysate10.1016/j.procbio.2007.08.011 ↗
52. Angiotensin-converting enzyme (ace) inhibitory activities of protein hydrolysates from Indonesian sea cucumbers10.1007/s10989-020-10035-5 ↗
53. Anti-inflammatory peptides and metabolomics-driven biomarkers discovery from sea cucumber protein hydrolysates10.1111/1750-3841.15834 ↗
54. Sun, S., Xu, X., Sun, X., Zhang, X., Chen, X., and Xu, N. (2019). Preparation and identification of ACE inhibitory peptides from the marine Macroalga Ulva intestinalis. Mar. Drugs, 17.10.3390/md17030179 ↗
55. Enzyme-induced gelation of extensively hydrolyzed whey proteins by alcalase: Peptide identification and determination of Enzyme Specificity10.1021/jf026242v ↗
56. Angiotensin Converting Enzyme Inhibitory Peptides Derived from Food Proteins: Biochemistry, Bioactivity and Production10.2174/138161207780363068 ↗
57. Characterization and storage stability of spray dried soy-rapeseed lecithin/trehalose liposomes loaded with a tilapia viscera hydrolysate
58. The preferential use of a soy-rapeseed lecithin blend for the liposomal encapsulation of a tilapia viscera hydrolysate
59. Effect of casein on pure lecithin liposome: Mixed biomacromolecular system for providing superior stabilization to hydrophobic molecules10.1016/j.colsurfb.2019.04.038 ↗
60. Ultrasonication for production of nanoliposomes with encapsulated soy protein concentrate hydrolysate: Process optimization, vesicle characteristics and in vitro digestion10.1016/j.fochx.2022.100370 ↗
61. Chitosan/sodium tripolyphosphate nanoparticles as efficient vehicles for antioxidant peptidic fraction from common kilka10.1016/j.ijbiomac.2018.01.023 ↗
62. Lima, K., Mauricio, C., Alemán, A., López-Caballero, M.E., Gómez-Guillén, M.C., Montero, M.P., and Prentice, C. (2021). Characterization, bioactivity and application of chitosan-based nanoparticles in a food emulsion model. Polymers, 13.10.3390/polym13193331 ↗
63. Potencial zeta en la determinación de carga superficial de liposomas
64. Chitosan nanoparticle as protein delivery carrier—Systematic examination of fabrication conditions for efficient loading and release10.1016/j.colsurfb.2007.04.009 ↗
65. Formulated chitosan-sodium tripolyphosphate nanoparticles for co-encapsulation of ellagic acid and anti-inflammatory peptide: Characterization, stability and anti-inflammatory activity10.1002/jsfa.12521 ↗
66. Liofilización de nanopartículas complejas de polielectrolitos: Efecto de la composición de nanopartículas y selección de crioprotectores10.1016/j.ijpharm.2018.09.035 ↗
67. Protein aggregation, water binding and thermal gelation of salt-ground hake muscle in the presence of wet and dried soy phosphatidylcholine liposomes10.1016/j.foodhyd.2018.04.025 ↗
68. Effect of simulated gastrointestinal digestion in vitro on the antioxidant activity, molecular weight and microstructure of polysaccharides from a tropical sea cucumber (Holothuria leucospilota)10.1016/j.foodhyd.2018.11.040 ↗
69. Encapsulation of oyster protein hydrolysates in nanoliposomes: Vesicle characteristics, storage stability, in vitro release, and gastrointestinal digestion10.1111/1750-3841.15606 ↗
70. Alemán, A., Marín-Peñalver, D., Fernández de Palencia, P., Gómez-Guillén, M.C., and Montero, M.P. (2022). Anti-Inflammatory properties, bioaccessibility and intestinal absorption of sea fennel (Crithmum maritimum) extract encapsulated in soy phosphatidylcholine liposomes. Nutriens, 14.10.3390/nu14010210 ↗
71. Release of multifunctiona l peptides by gastrointestinal digestion of sea cucumber (Isostichopus badionotus)10.1016/j.jff.2013.01.036 ↗
72. Effect of polymer molecular weight on chitosan-protein interaction10.1016/j.colsurfb.2014.11.037 ↗
73. Characterization of the interactions between chitosan/whey protein at different conditions10.1590/fst.29217 ↗
74. Improving resveratrol bioaccessibility using biopolymer nanoparticles and complexes: Impact of protein-carbohydrate Maillard conjugation10.1021/acs.jafc.5b00777 ↗
75. Improving carvacrol bioaccessibility using core-shell carrier-systems under simulated gastrointestinal digestion10.1016/j.foodchem.2021.129505 ↗
76. Preparation, characterization, and potential application of chitosan, chitosan derivatives, and chitosan metal nanoparticles in pharmaceutical drug delivery10.2147/DDDT.S99651 ↗
77. Mackie, A.R., Goycoolea, F.M., Menchicchi, B., Caramella, C.M., Saporito, F., Lee, S., Stephansen, K., Chronakis, I.S., Hiorth, M., and Adamczak, M. (2017). Innovative methods and applications in mucoadhesion research. Macromol. Biosci., 17.10.1002/mabi.201600534 ↗
78. Characterization of chitosan hydrochloride–mucin interaction by means of viscosimetric and turbidimetric measurements10.1016/S0928-0987(00)00065-8 ↗
79. Ways, T.M.M., Lau, W.M., and Khutoryanskiy, V.V. (2018). Chitosan and its derivatives for application in mucoadhesive drug delivery systems. Polymers, 10.10.3390/polym10030267 ↗
80. Why is chitosan mucoadhesive?10.1021/bm800276d ↗
81. A simple rheological method for the in vitro assessment of mucin-polymer bioadhesive bond strength10.1023/A:1015812615635 ↗
82. Comparison of different in vitro and ex vivo methods to evaluate mucoadhesion of glycol-palmitoyl chitosan micelles10.1016/S1773-2247(10)50073-X ↗
83. Evaluation of the mucoadhesive properties of chitosan nanoparticles prepared using different chitosan to tripolyphosphate (CS:TPP) ratios10.1016/j.ijbiomac.2018.09.185 ↗