In luence o Ala ia esculen a powde on senso y accep ance, nu i ional
aspec s, and s aling o whea b ead
Leona d Mihaly Cozmu a
a,*
, Anca Pe e
a
, Camelia Nicula
a
, Alexand a Ui a asan
a
,
Zsol Szak´
acs
a
, Sonia Sca i
b,c,d
, Maha Moussa
b,d
, Se ena Mi a a
b,c
, Paula Faja do Be n´
a dez
e
,
Sa a Baldassa i
, Giuliana D a a
, Anca Mihaly Cozmu a
a
a
Technical Uni e si y o Cluj Napoca, No h Uni e si y Cen e o Baia Ma e, Vic o iei S . 76, Baia Ma e, Romania
b
Depa men o Ea h, En i onmen and Li e Sciences, Uni e si y o Geno a, Co so Eu opa 26, 16132, Geno a, I aly
c
In e -Uni e si y Cen e o he P omo ion o he 3Rs P inciples in Teaching & Resea ch, 10129 Tu in, I aly
d
Na ional Biodi e si y Fu u e Cen e (NBFC), 90133 Pale mo, I aly
e
Bio echnology and Heal h Depa men , ANFACO-CYTMA, Colegio Uni e si a io, Vigo, Spain
Depa men o Pha macy, Uni e si y o Geno a, 16148, Geno a, I aly
ARTICLE INFO
Keywo ds:
Ala ia esculen a b ead
Glycemic index
B ead s aling
Func ional b ead
ABSTRACT
This s udy in es iga ed he nu i ional, senso y, physiological, and shel -li e impac s o inco po a ing Ala ia
esculen a seaweed powde in o whea b ead. S uc u al analysis showed ha high le els (7.5–10 %) educed
po osi y and olume, whe eas mode a e inclusion (2.5–5 %) main ained a o able c umb p ope ies. Senso y
e alua ion e ealed o e all accep abili y sco es o 8.15 o con ol b ead (B-C) and 8.12 o b ead wi h 5 %
A. esculen a (B-5A). P oxima e composi ion analysis showed inc eases in p o ein (1.26- old), ibe (1.47- old),
and ash (5.63- old) in B-5A, while ene gy alue and ca bohyd a e con en s dec eased (1.48- and 1.26- old,
espec i ely). In i o assays using THP-1 mac ophages and Caco-2 cells con i med he absence o cy o oxici y
up o 200 µg/mL, wi h sligh iabili y enhancemen s. In lamma o y gene exp ession (IL6, IL8, MCP1, TNF
α
)
emained unchanged, wi h a modes IL6 educ ion in Ala ia- ea ed mac ophages. In i o glycemic es ing
showed mode a ed pos p andial peaks o B-5A and B-C (≈130 mg/dL a 30 min) compa ed wi h glucose (146
mg/dL a 15 min), while B-5A main ained a mo e s able pla eau be ween 90–120 min, sugges ing p olonged
sa ie y. Du ing s o age, B-5A exhibi ed supe io mois u e e en ion (28.12 % s. 22.01 %), slowe s a ch
e og ada ion (44.76 %→47.95 % s. 45.27 %→50.29 %), and a mo e o de ed p o ein s uc u e, wi h g ea e
inc eases in
α
-helix (+14.13 %) and educ ions in andom coil (–12.40 %). P o ein su ace hyd ophobici y
e ol ed mo e g adually in B-5A (con ac angle dec ease 40.48◦→35.51◦) han in B-C (66.86◦→50.92◦), e lec ing
enhanced wa e -binding. Collec i ely, hese di e ences indica e slowe s aling and ex ended shel li e. O e all, 5
% A. esculen a o i ica ion enhances nu i ional quali y, delays s aling, imp o es shel li e, and main ains con-
sume accep abili y wi hou ad e se cy o oxic o in lamma o y e ec s.
1. In oduc ion
Whi e b ead, a s aple in a ious die s wo ldwide, has long been
c i icized o i s limi ed nu i ional alue and i s endency o s ale
apidly. While i emains a key ood p oduc in many cul u es due o i s
con enience and a o dabili y, i s d awbacks ha e p omp ed g owing
conce n ega ding i s ole in human heal h. These conce ns p ima ily
ocus on i s composi ion, which is based on e ined whea lou , esul ing
in a lack o essen ial nu ien s compa ed o whole-g ain b ead (Williams,
2012). A key issue wi h whi e b ead is i s high glycemic index, which is
associa ed wi h a apid inc ease in blood suga le els upon consump ion
and a highe isk o de eloping condi ions such as ype 2 diabe es,
ca dio ascula diseases, and obesi y (Lambadia i e al., 2020). Ano he
signi ican issue wi h whi e b ead is i s p one o apid s aling, a cascade
Abb e ia ions: AA sco e, amino acids sco e; B-C, con ol b ead; B-2.5A, b ead con aining 2.5% Ala ia esculen a epo ed o he weigh o lou in con ol b ead; B-
5A, b ead con aining 5% Ala ia esculen a epo ed o he weigh o lou in con ol b ead; B-7.5A, b ead con aining 7.5% Ala ia esculen a epo ed o he weigh o
lou in con ol b ead; B-10A, b ead con aining 10% Ala ia esculen a epo ed o he weigh o lou in con ol b ead.
* Co esponding au ho .
E-mail add ess: [email p o ec ed] (L. Mihaly Cozmu a).
Con en s lis s a ailable a ScienceDi ec
Fu u e Foods
jou nal homepage: www.else ie .com/loca e/ u o
h ps://doi.o g/10.1016/j. u o.2025.100805
Recei ed 2 Sep embe 2025; Recei ed in e ised o m 21 Oc obe 2025; Accep ed 21 Oc obe 2025
Fu u e Foods 12 (2025) 100805
A ailable online 24 Oc obe 2025
2666-8335/© 2025 The Au ho (s). Published by Else ie B.V. This is an open access a icle unde he CC BY-NC-ND license (
h p://c ea i ecommons.o g/licenses/by-
nc-nd/4.0/ ).
o p ocesses ha occu immedia ely a e baking (Amigo e al., 2021).
The s aling p ocess is d i en by he e og ada ion o s a ch, a phe-
nomenon in which he amylopec in molecules in he s a ch o b ead
c ys allize, leading o a loss o mois u e and a subsequen de e io a ion
in he ex u e and as e o he b ead (Willho , 2007). S ale limi s he
shel li e o whi e b ead, makes i less pala able, and mo e p one o
spoilage, hus con ibu ing o he economic and en i onmen al bu den
associa ed wi h ood was e (Ba ek e al., 2025). In ecen yea s, he e
has been g owing in e es in o i ying whi e b ead wi h unc ional in-
g edien s ha can enhance i s nu i ional p o ile and imp o e i s shel
li e. Seaweeds a e p omising ing edien s due o hei ich con en in
bioac i e compounds, including polysaccha ides, p o eins, i amins
(such as i amins A, C, and B12), essen ial mine als (such as iodine,
calcium, and magnesium), ca o enoids, phenolic compounds and sapo-
nins (Senadhee a e al., 2023). These bioac i e componen s o e a ange
o po en ial heal h bene i s, om suppo ing immune unc ion o
imp o ing gu heal h and educing he isk o ch onic diseases
(MacA ain e al., 2007).
Ala ia esculen a, a b own seaweed, is pa icula ly no able o i s high
con en o soluble ibe s, an ioxidan s, and mine als (Ly ou e al., 2021).
S udies ha e shown ha inco po a ing seaweed in o ood p oduc s can
enhance hei mine al con en , p o ide p ebio ic ibe s ha bene i gu
heal h, and impa an ioxidan p ope ies (Ma os e al., 2024). In b ead,
he addi ion o seaweed has been shown o imp o e he o e all nu i-
ional p o ile, inc ease ibe con en , and educe he glycemic index o
he inal p oduc (Qui al e al., 2022). Fu he mo e, seaweed has been
sugges ed o help mi iga e s aling due o i s hyd ophilic na u e, which
may assis in e aining mois u e and p olonging shel li e (Kılınç e al.,
2013). While A. esculen a o e s hese nu i ional and unc ional bene i s,
i can also accumula e hea y me als o excessi e iodine (Che y e al.,
2019), which may pose heal h isks i consumed in la ge amoun s o
o e p olonged pe iods. The e o e, i s elemen al composi ion should be
moni o ed o ensu e sa e y when used in ood applica ions.
Despi e he po en ial bene i s o o i ying b ead wi h seaweed, he e
is a no iceable gap in he scien i ic li e a u e ega ding he nu i ional
alue o b ead con aining A. esculen a. While s udies ha e examined he
inco po a ion o seaweed in o a ious ood p oduc s (Roohinejad e al.,
2017), ew ha e speci ically ocused on A. esculen a and i s e ec s when
used in b ead o mula ions. Mo eo e , he impac o A. esculen a on he
s aling p ocess o whi e b ead emains unde explo ed. Resea ch on he
ela ionship be ween he p esence o A. esculen a and he mechanisms o
b ead s aling, such as s a ch e og ada ion and changes in p o ein
s uc u e, could p o ide aluable insigh s in o how A. esculen a in-
luences b ead ex u e and shel li e. This could po en ially lead o new
unc ional ood p oduc s ha combine imp o ed nu i ional quali y wi h
ex ended eshness.
In his ega d, his s udy aims o explo e he po en ial o o i ying
whi e b ead wi h A. esculen a o imp o e he nu i ional p o ile o b ead
and po en ially mi iga e he apid s aling p ocess.
2. Ma e ials and me hods
2.1. Raw ma e ials and eagen s
Whi e lou (Baneasa, Romania), d y yeas (Pakmaya, Romania), sal
(Sal om, Romania), sun lowe oil (Unisol, Romania) we e pu chased
om a local supe ma ke .
Ala ia esculen a powde wi h g anulome y <2 mm was supplied by
The Seaweed Company (I eland). The Technical Shee p o ided by he
supplie is p esen ed in Table S1 om he Supplemen a y Ma e ial..
Reagen s HNO
3
, HCl, NaOH, 2,2-diphenyl-1-pic ylhyd azyl (DPPH),
asco bic acid, ace one, Folin-Ciocal eu, Na
2
CO
3
, chlo o o m, gallic acid,
pepsin, ypsin, chymo ypsin, ichlo oace ic acid (TCA), 2 % ninhyd in
solu ion, and l-glycine s anda ds we e pu chased om Me ck & Co., Inc.
(Da ms ad , Ge many).
2.2. B ead p epa a ion
The o mula ions o b ead loa es we e de eloped by pa ially
subs i u ing e ined whea lou wi h A. esculen a powde a di e en
inco po a ion le els (2.5 %, 5.0 %, 7.5 %, and 10 % w/w ela ed o he
whea lou con en ). The con ol o mula ion (B-C) consis ed exclu-
si ely o 1000 g o whea lou , whe eas he expe imen al o mula ions
(B-2.5A, B-5A, B-7.5A, and B-10A) con ained 975, 950, 925, and 900 g
o whea lou , espec i ely, wi h o he addi ion o 25, 50, 75, and 100 g
o A. esculen a powde , espec i ely. The amoun s o d y yeas (14 g) and
sun lowe oil (30 g) we e held cons an ac oss all o mula ions. The sal
concen a ion, howe e , was g adually educed om 15 g in con ol and
2.5 % subs i u ion b eads o 13 g, 12 g, and 10 g in he 5 %, 7.5 %, and
10 % o mula ions, espec i ely. This adjus men was based on p e-
limina y senso y e alua ions aimed a mi iga ing excessi e sal iness
a ibu able o he inhe en mine al con en o A. esculen a. The wa e
addi ion a ied wi h subs i u ion le el, anging om 600 mL in he
con ol dough o 640 mL in he 10 % subs i u ion, wi h inc emen al
inc eases o 10 mL pe o mula ion. The op imal wa e weigh was
de e mined h ough p elimina y hyd a ion capaci y es s. The in-
g edien s we e mixed o 15 min using a Bio i a MB-1500 PRO (Cluj
Napoca, Romania) a 2200 pm un il homogeneous doughs we e
o med. The doughs we e hen p oo ed o 1 hou in a e men a ion
chambe se o 30 ◦C and 75 % ela i e humidi y (Ma ina, Bucha es ,
Romania). The doughs we e shaped in o 750 g loa es by hand and
placed in baking ays (15 cm ×8 cm ×6 cm). They we e allowed o ise
o an addi ional 30 min a 30 ◦C and 75 % ela i e humidi y. The loa es
we e baked in a p ehea ed o en o 15 min a 200 ◦C, ollowed by 75 min
a 180 ◦C. A e baking, he loa es we e emo ed om he ays and
allowed o cool o 30 min a 25 ◦C be o e unde going senso y
e alua ion.
2.3. Appea ance o whole b ead and hei c oss-sec ions
Pic u es o eshly p epa ed b eads and selec ed b ead samples
du ing s aling we e cap u ed using a Canon 80D digi al single-lens e lex
came a (Canon Inc., Japan).
2.4. Senso y e alua ion
The senso y analysis o he b ead loa es was conduc ed a oom
empe a u e (22 ◦C) unde dayligh , wi h a g oup o 47 heal hy panel-
is s, o which 76.60 % we e emales and 23.40 % males, wi h ages be-
ween 18–50 yea s, who we e habi ual b ead consume s. The s udy was
app o ed by he E hics Commi ee o he Technical Uni e si y o Cluj-
Napoca (Romania, N . 594/10.04.2024) and adhe ed o he e hical
p inciples ou lined in he 1975 Decla a ion o Helsinki, as e ised in
2013. Be o e he senso y e alua ion, he pa icipan s we e in o med
abou he s udy’s pu pose and p o ided w i en consen . They we e also
ained o de elop a consensus on he desc ip i e ocabula y o he
b ead samples. The samples o each b ead o mula ion, cu in o 10×5 cm
pieces, we e coded, andomly placed on plas ic pla es, and p esen ed o
panelis s in a sequen ial monadic es ing o ma using a comple e block
design. In addi ion, panelis s we e gi en whole loa es o b ead and hei
c oss-sec ions. They we e asked o comple e a ques ionnai e in which
hey a ed he a ibu es o appea ance, colo , c us s eng h, c umb
s uc u e, elas ici y, olume, as e, smell, and o e all accep abili y using
a nine-poin hedonic scale, whe e 1-ex emely unpleasan ; 2-s ongly
unpleasan ; 3-unpleasan ; 4-sligh ly unpleasan ; 5-neu al; 6-sligh ly
pleasan ; 7-pleasan ; 8-s ongly pleasan ; 9-ex emely pleasan . To p e-
en he ca yo e e ec , pa icipan s insed hei mou hs wi h oom
empe a u e (25 ◦C) wa e be ween as ing each b ead o mula ion. The
o al sco e o each a ibu e was calcula ed as he a e age o he indi-
idual sco es. The A. esculen a-based b ead wi h he highes o e all
accep abili y sco e, as well as he con ol b ead, we e selec ed o u he
in es iga ions.
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
2
2.5. P oxima e analysis
P oxima e analysis o selec ed b ead loa es in e ms o mois u e
(AOAC 925.10, 1925), p o ein (AOAC 945.18-B, 1995, wi h a 6.25
ni ogen- o-p o ein con e sion ac o ), a (AOAC 920.39.C, 1995), ash
(AOAC 936.07, 2013), and ibe (Commission Regula ion No.
152/2009) was conduc ed acco ding o s anda dized me hods. The
ca bohyd a e con en was calcula ed using he equa ion:
Ca bohyd a es(g/100g)=100− − (Mois u e+P o ein+Fa +Ash+Fibe )
(1)
Conside ing he ene gy supplied by ca bohyd a es, p o eins, ibe ,
and a , he ene gy p o ided by B-C and B-5A was calcula ed using he
ollowing o mula:
2.6. Chlo ophylls and ca o enoids con en
Chlo ophylls and ca o enoids ex ac ion was pe o med in ace one a
a b ead- o-ace one a io o 10:1 (w: ). A e 1 min, he solu ion was
cen i uged o 15 min a 6000 x g. A olume o 15
μ
L o he eshly
p epa ed ex ac was dilu ed wi h 150
μ
L o ace one, and he abso bance
was spec opho ome ic measu ed (Pe kinElme Aanalys 800, SUA) a
662 nm, 645 nm, 470 nm, 653 nm, and 654 nm (Hyns o a e al., 2018;
Lich en hale and Buschmann, 2001). The esul s we e calcula ed using
Eqs. (3)–(8), and exp essed as
μ
g/100 g o d y b ead:
Ca =11.24∗A662 −2.04∗A645 (3)
Cb=20.13∗A645 −4.19 ∗A662 (4)
Ca +b=7.05∗A662 +18.09 ∗A645 (5)
Cx+c= (1000∗A470 −1.90 ∗Ca −63.14 ∗Cb)/214 (6)
Ccc= (Ca+b) + (Cx +c)(7)
Cph= (321.3∗A653) − (208.4∗A654)(8)
whe e Ca is he concen a ion o chlo ophyll a,
μ
g/mL; Cb is he con-
cen a ion o chlo ophyll b,
μ
g/mL; Ca+b is he concen a ion o o al
chlo ophylls,
μ
g/mL; Cx+c is he concen a ion o o al ca o enoids,
μ
g/
mL; Ccc is he concen a ion o o al chlo ophyll and ca o enoids,
μ
g/
mL; Cph is he concen a ion o o al pheophy ins,
μ
g/mL.
2.7. Cell cul u es and A. esculen a b ead ex ac s used in he in i o es s
Fo he sa e y assessmen o human consump ion o A. esculen a
based b ead, he po en ial oxici y o he B-5A was e alua ed in i o
using se e al physiologically ele an cellula models simula ing he
human gas oin es inal ac . In his ega d, he human in es inal Caco-2
cell line unde goes a spon aneous di e en ia ion in o a pola ised
cellula monolaye wi h unc ional ea u es o small in es inal en e -
ocy es, ul ima ely mimicking he human in es inal epi helial ba ie
(Fe uzza e al., 2012; Na oli e al., 2012). Simila ly, THP-1-de i ed
mac ophages ep esen a aluable in i o model o ob ain a mo e
in-dep h unde s anding o he in lamma o y p ocesses ca ied ou by
esiden in es inal mac ophages in he gas oin es inal ac
(Phuangbubpha e al., 2023). As a esul , bo h cellula models ha e been
ex ensi ely used o oxicological s udies (Susewind e al., 2016; Hemp
e al., 2020; Pole e al., 2020; Song e al., 2024; Gio dani e al., 2025).
The e o e, he cy o oxic po en ial o B-5A was e alua ed on
THP-1-de i ed M0 mac ophages, on undi e en ia ed Caco-2 cells and
on en e ocy e-like Caco-2-de i ed monolaye s a di e en ime poin s (i.
e., 24 and 72 h) using he MTT es . B-C was employed as a nega i e
con ol.
The human colon epi helial ca cinoma cell line Caco-2 was ob ained
om he In e lab Cell Line Collec ion (IRCCS Ospedale Policlinico San
Ma ino, Genoa, I aly) and cul u ed in high-glucose Dulbecco’s modi ied
Eagle medium (DMEM) (Biowes , Nuaill´
e, F ance) supplemen ed wi h
10 % FBS (Biowes ), 4 mM l-glu amine (Co ning Inc., NY, USA), 1 %
non-essen ial amino acids (Co ning Inc.), and penicillin/s ep omycin
(Co ning Inc.) as an ibio ics. Fo expe imen al p ocedu es, Caco-2 cells
we e used ei he as undi e en ia ed cells o as di e en ia ed en e ocy e-
like monolaye s o mimic he human in es inal epi helial ba ie . Di -
e en ia ion was achie ed by seeding Caco-2 cells a 300,000 cells/cm²
on T answell® PET inse s (12 mm diame e , 0.4
μ
m po e size) (VWR,
Milan, I aly) wi h comple e medium in bo h he apical and basola e al
compa men s. A e he o ma ion o a con luen monolaye , Caco-2
cells we e cul u ed in comple e medium o 21 days, wi h media
changes pe o med h ee imes a week (Fe uzza e al., 2012; Na oli
e al., 2012) o suppo hei di e en ia ion. Bo h cell lines we e
cul u ed a 37 ◦C in a humidi ied a mosphe e con aining 5 % CO₂.
The human monocy ic cell line THP-1 was sou ced om he Ame -
ican Type Cul u e Collec ion (LGC S anda ds s l, Milan, I aly) and
main ained in RPMI-1640 medium supplemen ed wi h 2 mM l-glu a-
mine (Eu oclone, Milan, I aly) and 10 % e al bo ine se um (FBS)
(Eu oclone). Fo expe imen al pu poses, THP-1 cells we e di e en ia ed
in o M0 mac ophages by he addi ion o 30 ng/mL pho bol-12-my is a e
13-ace a e (PMA) (Pep oTech EC, London, UK) o he cul u e medium
o 24 hou s (Di Giuseppe e al., 2022).
To p epa e he wa e -soluble b ead (WSB) ex ac s o he biological
es s, 10 g o d ied b ead samples (B-5A and B-C) we e dissol ed in 80
mL o deionized wa e and homogenised o 1 min wi h an ULTRA-
TURRAX® T25 basic (IKA-We ke GmbH, S au en, Ge many) be o e
sonica ion o 1 min a 100 Hz wi h an UP100H Ul asonic P ocesso
(Hielsche Ul asonics GmbH, Tel ow, Ge many). A e agi a ing o 30
min a oom empe a u e wi h a magne ic s i e , he WSB ex ac s we e
cen i uged a 4000 pm o 45 min and passed h ough a il e pape o
emo e esidual sun lowe oil om he supe na an . Once collec ed, he
ex ac s we e cen i uged a 14,000 pm o 10 min be o e he supe -
na an was s e ilized using a 0.22
μ
m sy inge il e (VWR®, Milan, I aly).
Fo he in i o oxicological expe imen s, each solu ion was dilu ed
di ec ly in he cul u e media o ob ain he di e en inal concen a ions
as epo ed below.
2.8. Toxici y assessmen wi h MTT es
The po en ial cy o oxici y o B-5A WSB was assessed a 24 and 72 h
in THP-1-M0 mac ophages and in Caco-2 undi e en ia ed cells. Fo bo h
ime-poin s, Caco-2 cells we e seeded a 15,000 cells/well in 96-well
pla es, while THP-1 monocy es we e pla ed a 50,000 cells/well and
induced o di e en ia e by ea men wi h PMA. Mo eo e , he po en ial
oxici y o B-5A WSB was also e alua ed in Caco-2 en e ocy e-like
monolaye s, di e en ia ed as desc ibed abo e. Fo all expe imen s, B-5A
and B-C WSB we e added o each well (200,100 and 50
μ
g/mL inal
concen a ions), and he pla es we e incuba ed a 37 ◦C o 24/72 h.
Ene gy alue(kcal/100g)= 4×Ca bohyd a e(%)+4×P o ein(%)+2×Fibe (%)+9×Fa (%)(2)
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
3
Then, cell iabili y was e alua ed by he MTT assay (0.5 mg/mL inal
concen a ion) as epo ed by Pozzolini e al. (2016). Da a a e he means
±SD o wo independen expe imen s in which each condi ion was
es ed eigh imes and esul s a e exp essed as pe cen age o cell iabili y
espec o un ea ed con ol cells (C).
2.9. In lamma o y cy okine exp ession by qPCR
To assess whe he consump ion o B-5A could somehow elici in-
lamma o y esponses, he gene exp ession o se e al in lamma o y
media o s (i.e., IL-1β, IL-6, IL-8, MCP1, and TNF-
α
) was in es iga ed in
THP-1-M0 mac ophages ea ed wi h B-5A and B-C WSB and he esul s
we e compa ed o hose o LPS posi i e con ol, a well-known p o-in-
lamma o y s imulus. THP-1 cells we e pla ed a 500,000 cells/well in 6-
well pla es and, once di e en ia ed in o M0 mac ophages, cells we e
challenged wi h B-5A and B-C WSB (200
μ
g/mL inal concen a ion) o
24 h. Then, he gene exp ession p o ile o in lamma o y media o s,
in e leukin-1β (IL-1β, a.n. NM_000576.3), in e leukin-6 (IL-6, a.n.
NM_001318095.2), in e leukin-8 (IL-8, a.n. NM_000584.4), monocy e
chemoa ac an p o ein-1 (MCP1, NM_002982), and umou nec osis
ac o -alpha (TNF-
α
, a.n. NM_000594.4) was e alua ed by qPCR ela i e
o un ea ed con ol cells. Values we e no malised o GAPDH (a.n.
NM_002046) and HPRT-1 (a.n. NM_000194.3) mRNA exp ession. RNA
ex ac ion, cDNA e o ansc ip ion and qPCR analyses we e pe o med
as desc ibed in Gio dani e al. (2025). All p ime pai s a e epo ed in
Table S2.
2.10. In i o glycemic index
The in luence o consuming con ol b ead and Ala ia-b ead on
pos p andial blood glucose le els was assessed in ela ion o aw
glucose. The s udy was app o ed by he E hics Commi ee o he Tech-
nical Uni e si y o Cluj-Napoca, Romania (no. 594/10.04.2024). A o al
o 50 heal hy subjec s, aged 18 o 60 yea s, we e included in he s udy.
Indi iduals wi h heal h condi ions we e excluded. Be o e he expe i-
men , subjec s we e in o med abou he objec i e o he s udy and
p o ided hei in o med consen . The s udy was conduc ed ollowing a
minimum o 10 hou s o as ing, ac oss h ee di e en days, wi h a one-
day b eak in be ween. The pa icipan s consumed 54.36 g o con ol
b ead (B-C), 80.50 g o Ala ia-b ead (B-5A), and 26.05 g o aw glucose
(adjus ed o he ca bohyd a e con en in he b ead) and d ank 100 ml o
wa e wi hin 10 min. Blood glucose le els we e measu ed using an Accu-
Chek Ac i e Glucome e (Roche Diabe es Ca e, Inc., Basel, Swi ze land),
wi h samples aken om he inge ip o each subjec e e y 15 min o e
a wo-hou pe iod ollowing he inges ion o he b ead loa es and
glucose. The expe imen was epea ed h ee imes.
2.11. S aling o selec ed b ead ypes
Following cooling o 1 hou , he b ead loa es we e s o ed in clean
polye hylene bags a 25 ◦C and 48 % ela i e humidi y o 4 days. The
s aling p ocess o he selec ed b ead samples was examined wi h espec
o he e olu ion o appea ance, changes in mois u e con en , c us
s eng h, speci ic olume, dispe sion coe icien , colo , changes in p o-
ein and s a ch s uc u es.
2.11.1. Speci ic olume
The apeseed displacemen me hod was used o de e mine he spe-
ci ic olume (V) o b ead, calcula ed using Eq. (9):
V(cm3/g)= (m1+m2−−m3)∗100/(m1∗d),(9)
whe e m
1
is he weigh o he b ead sample, g; d is he densi y o
apeseeds, 1.02 g/cm³; m
2
is he weigh o he con aine illed wi h
apeseeds, g; m
3
is he weigh o he con aine wi h bo h he sample and
apeseeds, g.
2.11.2. C us s eng h
C us s eng h was assessed using a Wagne FDK10 o ce dial pene-
ome e (Wagne Ins umen s, USA) on a 15-poin scale a he mini-
mum se ing, and he a e age alues we e epo ed.
2.11.3. Dispe sion coe icien in b ead loa es
The b ead loa was cu ho izon ally a i s highes poin , c ea ing a
c oss-sec ional su ace. The heigh (H) and diame e (D) o he loa we e
measu ed a i e di e en posi ions using a calipe , and he H/D index
was calcula ed based on he a e age alues.
2.11.4. Image analysis
The image analysis, conduc ed using ImageJ so wa e (Schneide
e al., 2012), in es iga ed he e olu ion o b ead c umb s uc u e o e a
ou -day s aling pe iod h ough solidi y analysis. Solidi y, a shape
desc ip o used o quan i y he compac ness o po es, was calcula ed as
he a io be ween he a ea o a po e and i s con ex a ea. Solidi y alues
ange om 0 o 1, whe e a alue o 1 indica es a e y compac , smoo h
(solid-like) po e, and lowe alues co espond o mo e i egula and
open po es. The solidi y alues we e g ouped in o nine ca ego ies (0.6,
0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, and 1.0), and he numbe o po es
(dimensionless) alling wi hin each solidi y ange was plo ed o Days 1
o 4.
2.11.5. Re lec ance spec a
The specia ion o indi idual-colo ed compounds in ood p oduc s is a
ime-consuming p ocess ha incu s high cos s o bo h ins umen a ion
and eagen s. Re lec ance spec a ha e been e ec i ely employed o
p edic he concen a ions o p o ein, amylose, o al polyphenols, and
ibe in ice, as well as i s an ioxidan ac i i y (Xie e al., 2014). How-
e e , no li e a u e has been ound ega ding he use o e lec ance
spec a o iden i y op ically ac i e compound g oups and hei
co-pigmen a ion in b ead o i ied wi h seaweed. Such an app oach
could p o ide insigh s in o he senso y p ope ies o en iched b ead,
such as an ioxidan ac i i y, speci ic as es in e ms o as ingency and
bi e ness.
The colo changes du ing he s aling p ocess o con ol and Ala ia-
en iched b ead we e e alua ed h ough he analysis o e lec ance
spec a. Re lec ance spec a o he samples a a ious s ages o s aling
we e eco ded o e a wa eleng h ange o 360 o 780 nm, wi h 10
epe i ions a a 1 nm esolu ion. Da a acquisi ion was pe o med using a
UV/VIS Lambda 35 spec ome e (Pe kinElme , USA), coupled wi h a
Labsphe e RSA-PE-20 in eg a ing sphe e. The e lec ance spec a we e
decon olu ed in o se en disc e e wa eleng h bands co esponding o
de ined egions o he isible spec um: 380–449 nm (pu ple), 450–484
nm (blue), 485–499 nm (cyan), 500–564 nm (g een), 565–589 nm
(yellow), 590–624 nm (o ange), and 625–760 nm ( ed). These spec al
egions a e indica i e o op ically ac i e, colo -con ibu ing compounds
p esen in he b ead samples. Thei dis ibu ion p o ides aluable in-
o ma ion on pigmen composi ion as well as po en ial chemical al e -
a ions ela ed o o mula ion di e ences and s aling p ocess.
2.11.6. FTIR spec a
The FTIR abso bance spec a o he b ead samples we e eco ded
using a Pe kin Elme BX2 spec ome e (SUA) equipped wi h an ATR
Pike Mi acle diamond c ys al, co e ing a measu emen ange o
1550–6000 cm⁻¹. FTIR analysis was pe o med by placing he b ead
samples di ec ly on o he c ys al unde he same p essu e. Backg ound
spec a we e eco ded p io o sample measu emen . The samples we e
analyzed wi hou any p io p epa a ion o p ese e he na i e s a ch and
p o ein s uc u es, as p ocessing could induce s uc u al al e a ions.
Spec a we e collec ed a a esolu ion o 4 cm⁻¹ by accumula ing 30
epe i ions. S uc u al changes in s a ch and he seconda y s uc u e o
p o eins (amide I band) we e analyzed in he anges o 4000–6000 cm⁻¹
and 1550–1700 cm⁻¹, espec i ely. To achie e his, FTIR spec a we e
p ocessed acco ding o he me hod p oposed by Mihaly Cozmu a e al.
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
4
(2025) h ough no maliza ion and decon olu ion, using he Sa i z-
ky–Golay coe icien s o a hi d-o de polynomial wi h a 25-poin
window, assuming a Gaussian dis ibu ion o he minima o he
second-o de de i a i e. The decon olu ed cu es we e analyzed in
e ms o he ela i e c ys allini y deg ee o s a ch (1047 cm⁻¹ and 995
cm⁻¹ o he c ys alline o m, and 1020 cm⁻¹ o he amo phous o m)
(Wang e al., 2020). Fo p o ein s uc u e, he a io o seconda y
s uc u al ea u es was assessed, including
α
-helix (dense o de ed
s uc u e, 1663–1646 cm⁻¹), β-shee (o de ed s uc u e, 1636–1615
cm⁻¹, 1700–1682 cm⁻¹), β- u n ( ela i ely ex ended o de ed s uc u e,
1681–1664 cm⁻¹), and andom coil (diso de ed s uc u e, 1645–1637
cm⁻¹) (Zhao e al., 2021).
2.11.7. Su ace hyd ophobici y o b ead p o ein isola e
P o ein ex ac ion was pe o med ollowing he me hod desc ibed by
Joshi e al. (2011). B ead samples we e mixed wi h dis illed wa e a a
a io o 1:15 (w: ), and he pH o he solu ion was adjus ed o 9.5 using a
1.0 M NaOH solu ion. The mix u e was homogenized o one hou a
oom empe a u e while s i ing a 750 pm. A e emo al o he coa se
solid phase, he solu ion was cen i uged a 6000 x g o 30 min. The pH
o he supe na an was hen adjus ed o 4.5 using a 1.0 M HCl solu ion.
P o ein p ecipi a ion was allowed o occu a oom empe a u e (25 ◦C)
o 5 hou s. Following a second cen i uga ion unde he same condi-
ions, he liquid phase was disca ded, and he p o ein isola e was
collec ed. I was washed wi h dis illed wa e o emo e soluble com-
ponen s, neu alized o pH 7 wi h 1.0 M HCl, and he p o ein suspension
was decan ed and il e ed. The p o ein isola e was hen d ied a 35 ◦C o
24 hou s.
The e olu ion o su ace hyd ophobici y o he p o ein isola es du -
ing b ead s aling was discussed based on he con ac angle o a wa e
d op, measu ed using he sessile d op me hod (Zhao e al., 2021). The
p o ein isola e was sp ead on a glass slide, and a 5
μ
L d op o dis illed
wa e was placed on i s su ace. Images we e cap u ed using a Canon EF
100 mm /2.8 L Mac o IS USM lens, se o i s highes magni ica ion (1x)
and a high dep h o ield ( /13). Focus was adjus ed by posi ioning he
sample on a mic oscope sample holde . No addi ional whi e balance o
image pos -p ocessing was applied, aside om he au oma ic se ings
applied by he came a. The con ac angles we e measu ed using ImageJ
(Schneide e al., 2012) so wa e. The con ac angle was eco ded on
bo h sides o each d op, wi h measu emen s aken wi hin 15 s o p e en
wa e adso p ion by he pelle .
2.12. S a is ical analysis
The expe imen s we e conduc ed in iplica es, and he esul s a e
p esen ed as he mean ±s anda d de ia ion. Con ac angle and c us
s eng h measu emen s we e aken om a leas 10 eplica es, and he
esul s a e also exp essed as mean ±s anda d de ia ion. To assess sig-
ni ican di e ences be ween g oup means, one-way ANOVA ollowed by
Tukey’s es (G aphPad So wa e, Inc., San Diego, CA, USA), esul s wi h
p alues <0.01 o p <0.05 we e conside ed signi ican .
P incipal Componen Analysis (PCA) was pe o med using S a is ica
7.0 (S a So Inc., Tulsa, USA) o highligh he unde lying pa e ns and
ela ionships be ween he b ead o mula ion and s aling pa ame e s,
enabling he iden i ica ion o key ac o s d i ing a iabili y and hei
con ibu ions o he o e all da a s uc u e.
3. Resul s and discussion
3.1. Appea ance o b ead a ie ies
A comp ehensi e analysis o he s uc u al and ex u al a ibu es o
he o mula ed b eads is p esen ed in Table 1. Based on hese indings, i
can be concluded ha he inco po a ion o low concen a ions (2.5 %–5
%) o Ala ia esculen a powde esul s in b eads wi h a a o able balance
be ween s uc u al in eg i y and nu i ional enhancemen . Highe
inclusion le els (7.5 %–10 %) al e he c umb ex u e, educing po osi y
and making he b ead dense . Al hough A. esculen a con ibu es o a
da ke c umb colo and may enhance he nu i ional alue o b ead,
excessi e inco po a ion nega i ely impac s key quali y pa ame e s such
as ex u e, elas ici y, and speci ic olume, ac o s ha a e likely o in-
luence consume accep abili y.
3.2. Senso y s udy
The esul s o he senso y e alua ion o b ead samples con aining
a ying pe cen ages o Ala ia esculen a powde (B-C, B-2.5A, B-5A, B-
7.5A, B-10A) a e p esen ed in Figu e S1. Conce ning appea ance, he
con ol b ead (B-C) ecei ed he highes sco e, indica ing ha i s isual
cha ac e is ics we e mos appealing o consume s. As A. esculen a
powde was inco po a ed, a g adual decline in appea ance sco es was
no ed, pa icula ly in B-7.5A and B-10A. The da ke colo a ion and
po en ial speckling due o he seaweed powde may ha e con ibu ed o
lowe accep abili y. Howe e , B-2.5A and B-5A main ained a o able
isual appeal, sugges ing ha mode a e le els o A. esculen a powde do
no signi ican ly impac he appea ance o he b ead. A oma sco es
declined as he pe cen age o Ala ia esculen a powde inc eased. B-C had
he highes a oma accep abili y, whe eas B-10A ecei ed he lowes
sco es. The in oduc ion o seaweed powde impa ed ma ine-like no es,
which may no ha e me he consume expec a ions o b ead. B-2.5A
and B-5A main ained a mo e neu al a oma p o ile, sugges ing ha a
hese concen a ions, he impac on a oma is minimal. The ex u e
sco es ollowed a simila downwa d end in b ead wi h inc easing
A. esculen a con en . While B-C had he mos a o able ex u e a ing, B-
2.5A and B-5A emained wi hin an accep able ange. Howe e , B-7.5A
and B-10A showed signi ican declines, likely due o changes in c umb
s uc u e and mois u e e en ion caused by he seaweed powde . Highe
concen a ions o A. esculen a may ha e a ec ed he elas ici y and
so ness o he b ead, making i less pala able. Tas e sco es e lec ed a
decline in consume p e e ence wi h inc easing A. esculen a con en .
Also in his case, B-C had he highes a ing, while B-10A exhibi ed he
lowes . The p esence o umami and ma ine la o s in highe concen-
a ions may ha e con ibu ed o he lowe sco es. Howe e , B-2.5A and
B-5A emained accep able, indica ing ha small amoun s o A. esculen a
powde can be inco po a ed wi hou d as ically al e ing he as e p o-
ile. The o e all accep abili y sco es e eal an in e es ing end. While
he con ol b ead, B-C, ecei ed he highes a ing (8.15), he B-5A
sample (8.12) closely ollowed, indica ing ha a 5 % A. esculen a powde
inclusion does no signi ican ly impac consume p e e ence. The B-2.5A
sample (7.74) was sligh ly lowe bu s ill wi hin an accep able ange.
These esul s sugges ha mode a e inco po a ion o A. esculen a powde
(up o 5 %) does no d as ically educe accep abili y and may e en be
compa able o he con ol. Howe e , highe inclusion le els (B-7.5A and
B-10A) in oduced s onge senso y changes ha led o lowe o e all
sco es. The senso y s udy indica es ha he inclusion o A. esculen a
powde a ec s consume accep abili y ac oss mul iple senso y pa am-
e e s. Highe concen a ions (B-7.5A and B-10A) led o a decline in
p e e ence, while lowe le els (B-2.5A and B-5A) main ained an
accep able senso y p o ile. No ably, he B-5A sample exhibi ed an
o e all accep abili y sco e e y close o con ol, B-C, sugges ing ha i
may be a p omising o mula ion.
The senso y s udy conduc ed by J¨
onsson e al. (2024) on whea b ead
con aining 7.5 % Ala ia esculen a ( ela i e o whea lou ), in ol ing 49
consume s (45 % women and 55 % men), epo ed he ollowing sco es
o Ala ia-based b ead: 6.5 o appea ance, 5.9 o smell, 5.7 o as e,
6.2 o ex u e, and 5.9 o o e all. In compa ison, he co esponding
sco es o he con ol b ead we e 6.2, 6.2, 6.1, 6.5, and 6.3, espec i ely.
Apa om appea ance, whe e he b ead wi h 7 % Ala ia esculen a
ecei ed a highe sco e han he con ol b ead, he smell, as e, ex u e,
and o e all accep abili y we e a ed wi h lowe sco es. Fo he a ibu es
o appea ance, smell, as e, and ex u e, he sco es ecei ed by he b ead
con aining 7.5 % A. esculen a in ou s udy a e simila o hose epo ed
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
5
Table 1
S uc u al and ex u al cha ac e is ics o he in es iga ed b eads.
Fea u e Con ol B ead (B-C) B ead wi h 2.5% Ala ia esculen a (B-2.5A) B ead wi h 5% Ala ia esculen a (B-5A) B ead wi h 7.5% Ala ia esculen a (B-
7.5A)
B ead wi h 10% Ala ia esculen a (B-10A)
Colo C umb: B igh whi e C us : Golden-
b own
C umb: Ligh beige, mino g eenish in
C us : Golden-b own wi h sligh dullness
C umb: Da ke beige wi h g een speckles
C us : Ligh b own, sligh ly ough
C umb: B own-g een, uni o m da kening
C us : Thicke , oughe , da ke b own
C umb: Deep b own-g een, almos
g ayish C us : Da k, igid, c acked
C umb densi y Ligh , lu y,
well- isen
Sligh ly dense , s ill so No iceably dense , sligh ly educed
expansion
Much dense , signs o collapse Ex emely dense, c umbly, weak
in e nal s uc u e
Po osi y Highly po ous, ine, uni o m
dis ibu ion o po es
Sligh ly educed po osi y, small and e en
po es
Mode a e po osi y, smalle po es, sligh ly
compac ed
I egula po osi y, une en bubble
dis ibu ion
Low po osi y, la ge oids and collapse
a eas
Elas ici y &
sp ing
High elas ici y, ebounds well S ill e ains elas ici y, mino i mness
inc ease
Mode a e elas ici y, i me s uc u e Reduced elas ici y, c umb becomes igid Low elas ici y, c umb b eaks easily
Mois u e
e en ion
Balanced mois u e, so and esh Sligh ly d ie bu accep able No iceably i me , mino d yness Much lowe mois u e, i me and d ie
c umb
D y, b i le, lowe wa e e en ion
Glu en ne wo k
s eng h
S ong, well- o med glu en s ands
suppo ing s uc u e
Mos ly in ac , mino weakening Sligh glu en weakening, causing educed
ise
Signi ican glu en ne wo k weakening,
less ise & s uc u e
Weak glu en ne wo k, leads o
s uc u al collapse
Po es size and
dis ibu ion
Small o medium, e enly dis ibu ed Mos ly small and uni o m Smalle , nume ous, bu consis en I egula , la ge oids in some a eas La ge, inconsis en , collapse-p one ai
pocke s
C umb ex u e Smoo h, ligh ,
well-s uc u ed
Sligh ly i me bu smoo h Dense , somewha oughe Roughe c umb, ib ous mou h eel Ex emely ough, ib ous, b i le
C us hickness Thin, c isp c us wi h so in e io Sligh ly hicke c us , emains c isp Thicke c us , sligh ly i me No iceably hicke , ha de c us Ve y hick, igid c us , la ge c acks
isible
S uc u al
in eg i y
S ong, holds shape well when sliced Main ains shape, mino i mness inc ease Holds shape bu dense Some de o ma ion, weakened s uc u e Weak s uc u e, c umbles and collapses
when sliced
Loa expansion Well- isen, ai y, symme ical loa Good expansion, mino heigh educ ion Reduced heigh , i me c umb Clea loss o ise, sligh ly sunken S uc u al collapse, compac ed loa
A oma T adi ional whea a oma, b igh whi e
c umb
Mild seaweed hin s, sligh beige hue S onge ea hy a oma, isible g een lecks Mo e p onounced ea hy/seaweed scen ,
da ke one
S ong seaweed a oma, deep da k
c umb
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
6
by J¨
onsson e al. (2024) o b ead con aining 7 % A. esculen a. Howe e ,
o he o e all accep abili y a ibu e, ou s udy epo ed a sligh ly
highe sco e o 6.46 compa ed o 5.9 in he s udy o J¨
onsson e al.
(2024). The simila sco es a ed o he b ead con aining A. esculen a by
consume s om wo dis inc coun ies sugges ha i s senso y a ibu es,
such as appea ance, smell, as e, ex u e, and o e all accep abili y, we e
pe cei ed simila ly ac oss hese g oups. This could indica e ha he
inclusion o A. esculen a in b ead may o e a senso y p o ile ha is
accep able in di e en cul u al con ex s, p o iding ini ial e idence o
i s po en ial in b oade ma ke s.
Based on he esul s o he senso y s udy, he b ead con aining 5 %
A. esculen a powde (B-5A) and he b ead wi hou he seaweed powde
(B-C) we e selec ed o u he in es iga ion.
3.3. P oxima e composi ion
The p oxima e composi ion o he selec ed b eads is displayed in
Table 2. The mois u e con en o B-5A is signi ican ly highe han ha o
B-C, due o he wa e -binding p ope ies o A. esculen a (Benne e al.,
2023), which could esul in mois e b ead. Inc eased mois u e can
imp o e he so ness and mou h eel o he b ead; howe e , i may also
impac i s shel li e by p omo ing mic obial g ow h i no p ope ly
con olled. The p o ein con en in B-5A is signi ican ly highe compa ed
o he con ol b ead. The inco po a ion o A. esculen a, which is a
seaweed known o i s high p o ein con en (Table S1, Benne e al.,
2023), clea ly con ibu es o his inc ease. The highe p o ein con en
could also enhance he unc ional p ope ies o he b ead, such as
imp o ing i s s uc u e and ex u e. Bo h b eads exhibi low- a con en ,
wi h no signi ican di e ence be ween he alues. This sugges s ha he
addi ion o A. esculen a powde does no al e he a p o ile o he b ead,
e lec ing he in insic composi ion o he seaweed, which is no a sig-
ni ican sou ce o a (Table S1). A signi ican inc ease in ibe con en
was obse ed in B-5A.A. esculen a is known o i s high ibe con en
(Table S1), which con ibu es o imp o ed diges i e heal h and may
help egula e blood suga le els. The highe ibe con en in B-5A may
also enhance sa ie y, which could be bene icial o weigh managemen .
The ash con en in B-5A is signi ican ly highe han in B-C, indica ing a
highe mine al pe cen age.
Gi en he ypical ecommended daily in ake (RDI) o each nu ien
in adul s, in e ms o 114 g/day p o ein o men and 102 g/day p o ein
o women, 25 g/day ibe , 130 g/day ca bohyd a es, 70 g/day a , and
2000 kcal/day ene gy (EFSA, 2017) he con ibu ion o he selec ed
b eads o he RDI was calcula ed (Table 2). B-5A p o ides a highe
pe cen age o he daily p o ein equi emen compa ed o B-C, making i
a mo e p o ein- ich op ion. B-5A also o e s mo e ibe han B-C,
con ibu ing o a signi ican ly highe pe cen age o he daily ecom-
mended in ake, which could be pa icula ly bene icial o he diges i e
sys em. On he o he hand, he con ol b ead, B-C, p o ides a g ea e
pe cen age o he daily ca bohyd a e equi emen , while B-5A con ains
ewe ca bohyd a es, which may appeal o low ca bohyd a e die s. Bo h
b eads a e endowed wi h e y small amoun s o a ela i e o he daily
ecommended in ake (a ound 1 %). The ene ge ic alue o B-5A is lowe
han ha o he con ol b ead, e lec ing he educ ion in ca bohyd a e
con en . The lowe calo ic densi y o B-5A may be ad an ageous o
consume s aiming o educe calo ic in ake while bene i ing om highe
p o ein, ibe , and mine al con en . Howe e , his educ ion in ene gy
may also in luence he ex u e and mou h eel, po en ially making he
seaweed-based b ead dense . The di e ences be ween he in es iga ed
b eads highligh he po en ial bene i s o inco po a ing Ala ia esculen a
in o b ead p oduc s, pa icula ly o consume s seeking enhanced ibe ,
p o ein, and mine al in ake, while also educing ca bohyd a e and
calo ic consump ion.
3.4. Ci o oxici y s udy and in lamma o y cy okine exp ession
Fo consis ency wi h he composi ion o he luids ha he b ead
would encoun e in he diges i e ac i consumed, he aqueous ex ac s
(WSB) o he con ol b ead, B-C, and o he 5 % A. esculen a-en iched
b ead, B-5A, we e ob ained and hen es ed in in i o oxici y and in-
lamma o y esponse s udies. No signi ican di e ences we e obse ed
in he o al phenolic con en (~2.1
μ
g/mg ex ac ) and in he DPPH
an ioxidan sca enging ac i i y (~21 % a 2.5 mg/mL ex ac ) o he
wo B-C and B-5A b eads (wo k p o ocols a e p o ided in he Supple-
men a y Ma e ial), indica ing a simila composi ion o he compounds
ex ac ed in he wo b eads.
Conce ning he MTT cy o oxici y es s, in di e en ia ed THP-1 M0
mac ophages, a he 24-hou ime poin (Fig. 1A), all ex ac s, ega dless
o b ead ype o concen a ion, main ained cell iabili ies a o abo e
100 %, indica ing no ha m ul e ec s. A he highes concen a ion (200
µg/mL), a sligh inc ease in cell iabili y was obse ed o bo h b ead
ypes, wi h alues modes ly exceeding ha o he con ol, un ea ed
cells. A 100 and 50 µg/mL, iabili y emained s able, wi h WSB om
he B-5A Ala ia-en iched b ead consis en ly exhibi ing sligh ly highe
mean alues compa ed o con ol b ead ex ac s. O e all, no signi ican
cy o oxici y was de ec ed a e 24 hou s o exposu e. The p esence o
A. esculen a powde did no ad e sely a ec mac ophage iabili y and
may e en con e a modes iabili y-enhancing e ec a lowe concen-
a ions. A e 72 hou s o exposu e (Fig. 1B), a simila end o high cell
Table 2
P oxima e composi ion o con ol b ead and b ead con aining 5 % Ala ia esculen a powde .
Pa ame e RDI, g/day (EFSA,
2017)
Con ol b ead, B-C B ead wi h 5% Ala ia, B-5A
Amoun Con ibu ion o he RDI/100g o
b ead
Amoun Con ibu ion o he RDI /100g o
b ead
Mois u e, g/100 g- 37.25 ±0.60
b
- 41.32 ±0.70
a
-
P o ein, g/100 g
(d y sample)
114 o men
102 o women
10.12 ±0.04
b
8.88 % o men
9.92 % o women
12.73 ±0.08
a
11.17 % o men
12.48 % o women
Fa , g/100 g
(d y sample)
70 0.85 ±0.04
a
1.21 % 0.72 ±0.02
a
1.03 %
Fibe , g/100 g
(d y sample)
25 2.40 ±0.00
b
9.60 % 3.53 ±0.00
a
14.12 %
Ash, g/100 g
(d y sample)
- 1.66 ±0.01
b
- 9.34 ±0.03
a
-
Ca bohyd a es, g/100
(d y sample)
130 47.92 ±0.88
a
36.86 % 32.36 ±0.54
b
24.89 %
Ene ge ic alue, Kcal/100 g (d y
sample)
2000 243.81 ±
4.75
a
12.19 % 193.90 ±
3.70
b
9.70 %
RDI – ecommended daily in ake; The con ibu ions o he RDI we e calcula ed based on EFSA (2017) guidelines o adul males and emales aged 19–50 yea s.
Indi idual equi emen s may a y based on age, sex, heal h s a us, and physiological condi ions.
Da a a e p esen ed as mean ±s anda d de ia ion (n =3). Values wi h di e en le e s a and b in he same line a e signi ican ly di e en (p <0.05).
Values o p o ein, a , ibe , ash, ca bohyd a es and ene ge ic alue a e epo ed o he d y sample.
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
7
(cap ion on nex page)
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
8
iabili y pe sis ed. A sligh inc ease abo e con ol le els was again
obse ed o bo h ex ac s, B-C and B-5A WSB, pa icula ly a 200 µg/
mL, whe e he B-5A WSB appea ed o p omo e ma ginally g ea e
iabili y han he B-C. A 100 µg/mL and 50 µg/mL, cell iabili y
emained a high le el. While he di e ences we e mino , he B-5A WSB
consis en ly demons a ed a sligh ele a ion in iabili y ela i e o he
con ol.
Compa able ou comes we e ob ained in undi e en ia ed Caco-2 cells
(Fig. 1C and D). Ac oss all concen a ions es ed (up o 200 µg/mL), bo h
B-C and B-5A WSB we e non-cy o oxic. The inclusion o 5 % A. esculen a
powde did no nega i ely impac cell iabili y and, in some cases, may
ha e sligh ly imp o ed i , pa icula ly a he 24-hou ime poin
(Fig. 1C). Upon p olonged exposu e o 72 hou s (Fig. 1D), iabili y
ends emained s able o sligh ly inc eased, sugges ing no accumula ion
o cy o oxic e ec s o e ime. The minimal a iabili y be ween 24 h and
72 h da a u he suppo s he biocompa ibili y o bo h ex ac ypes.
These indings sugges ha he inco po a ion o Ala ia esculen a in o
b ead does no comp omise in es inal epi helial cell iabili y and may
help sus ain cellula homeos asis.
Simila ly, no cy o oxici y was obse ed in di e en ia ed Caco-2
en e ocy e-like monolaye s ollowing 24 and 72 hou s o ea men
wi h he ex ac s. A 24 hou s (Fig. 1E), cell iabili y emained consis-
en ly high ac oss all ex ac ypes, wi h no signi ican educ ions
obse ed. In e es ingly, a e 72 hou s (Fig. 1F), he B-5A WSB a 200
µg/mL appea ed o induce a sligh inc ease in me abolic ac i i y, wi h
iabili y app oxima ely 8.1 % highe han ha o con ol, un ea ed
cells. These indings con i m ha A. esculen a powde inclusion in b ead
does no comp omise en e ocy e-like Caco-2 cell wel a e e en a e
ex ended exposu e. No ably, while he B-C WSB alone may sligh ly
enhance cell p oli e a ion a highe concen a ions o e 72 hou s, he
p esence o A. esculen a in he B-5A b ead appea s o egula e his e ec ,
main aining cell iabili y nea baseline le els. This may sugges a
modula o y in luence o A. esculen a on cell p oli e a ion, wi hou
exe ing cy o oxic s ess.
Ou esul s align wi h he indings o O’Sulli an e al. (2011), who
epo ed ha me hanolic ex ac s o b own seaweeds (e.g., Ascophyllum
nodosum, Lamina ia hype bo ea, Pel e ia canalicula a, Fucus esiculosus,
and Fucus se a us) did no comp omise Caco-2 cell iabili y a concen-
a ions up o 2 mg/mL, as de e mined using he MTT assay. Also, he
s udy o Huwai e al. (2022) which indica ed ha ucoidan, an
algal-de i ed polysaccha ide, exp essed no cy o oxici y on THP-1 mac-
ophages, e en a 200 µg/mL o 24 h.
Conce ning he in lamma o y po en ial in i o s udy, as expec ed,
exposu e o lipopolysaccha ide (LPS) posi i e con ol esul ed in a sig-
ni ican ansc ip ional up egula ion o IL-6, IL-8, MCP-1, and TNF-
α
in
THP-1 mac ophages a e 24 hou s, wi h old inc eases o 2.8, 1.5, 1.8,
and 1.6, espec i ely, compa ed o un ea ed con ols (Fig. 1G). In
con as , ea men wi h ei he B-C o B-5A WSB did no elici signi ican
al e a ions in he exp ession o hese in lamma o y media o s. Howe e ,
a sligh educ ion in IL-6 mRNA le els was obse ed in mac ophages
ea ed wi h he B-5A WSB. Also, no signi ican di e ences in cy okine
exp ession we e de ec ed be ween he wo b ead WSB ea men s.
O e all, he in i o oxicological indings suppo he sa e y o
A. esculen a-en iched b ead, as nei he oxic no p o-in lamma o y e -
ec s we e obse ed in he examined in es inal (Caco-2) and immune
(THP-1 mac ophage) cell models. Bioac i e compounds commonly
ound in b own seaweeds, including ucoidan, phlo o annins, ucoxan-
hin, and s e ols, ha e been epo ed o modula e oxida i e s ess-
ela ed signaling pa hways, enhance cellula de ense mechanisms, and
exhibi he apeu ic po en ial in he managemen o in lamma ion-
associa ed diso de s (Begum e al., 2021). Speci ically, ucoidan ex-
ac s de i ed om species such as Unda ia pinna i ida, Fucus esiculosus,
Ascophyllum nodosum, Lamina ia japonica, and Mac ocys is py i e a ha e
been shown o supp ess he sec e ion o TNF-
α
, IL-1β, and IL-6 in human
pe iphe al blood mononuclea cells (PBMCs) and THP-1 mac ophages
ollowing LPS s imula ion (Ahmad e al., 2021). Fu he mo e, in
THP-1-de i ed mac ophages, ucoidan has been demons a ed o
a enua e he elease o p o-in lamma o y cy okines and inhibi cell
mig a ion (Li e al., 2017). Fu u e in es iga ions, by means o o gano-
ypic in i o models o in es inal in lamma ion, will help in highligh ing
he abo e-men ioned possible unc ional e ec s o A. esculen a en iched
unc ional oods such as he b ead designed in ou s udy.
3.5. In i o glycemic index
Fig. 2 shows he pos p andial blood glucose esponse wi hin 120 min
a e he consump ion o aw glucose, con ol b ead (B-C), and b ead
wi h 5 % Ala ia esculen a (B-5A), wi h s anda d de ia ions o alues
anging be ween 6.73 % and 36.80 %. The ANOVA analysis o esul s
(Table S3) did no indica e signi ican (p <0.05) di e ences due o high
da a a iabili y. S a is ically alida ed conclusions we e ob ained by
using he da a be ween he 25 % and 75 % qua iles co esponding o he
bes esul s in he middle ange o a ia ion, wi h s anda d de ia ions
anging be ween 1.64 % and 12.87 % (Table S4).
The p o ile o aw glucose indica es a apid inc ease, wi h he
highes blood glucose le el o 146.00 mg/dL eached a e 15 min. A e
peaking, blood glucose d opped sha ply up o 45 min o 105.6 mg/dL
and hen con inued declining s eadily, eaching nea ly baseline a 89.6
mg/dL a e 120 min. A mode a e ise in blood glucose was obse ed o
he con ol b ead, wi h a maximum o 130.5 mg/dL a e 30 min, ol-
lowed by a g adual decline o nea -baseline le el a 96.2 mg/dL a e
120 min. A simila end was also obse ed o A. esculen a-b ead, B-5A,
wi h no signi ican di e ences in he maximum alue o he ime
equi ed o each i compa ed o he con ol b ead. The decline was
g adual bu sligh ly slowe han in B-C, wi h inal blood glucose le el o
94.00 mg/dL, no signi ican ly di e en om ha o B-C. The compa -
a i e analysis o he e olu ion o blood glucose esponses indica es ha ,
a e he consump ion o aw glucose, he glucose peak in he blood is
eached as e (15 min) and a a highe le el (146.00 mg/dL) compa ed
o he consump ion o B-C and B-5A (30 min, 130.50 mg/dL and 133.80
mg/dL, espec i ely). In he case o B-5A, he e olu ion o blood glucose
le el was no signi ican ly di e en om ha o con ol b ead du ing he
90 min, bu i emained a a highe alue han B-C du ing he 90–120
min ange. I sugges s ha b ead o i ied wi h 5 % A. esculen a no only
con ols he magni ude o he pos p andial glycemic esponse ela i e o
aw glucose inges ion bu also in luences he empo al p o ile o blood
glucose le els compa ed o con ol b ead (B-C). While bo h b ead sam-
ples show a lowe glycemic peak han aw glucose, B-5A main ained a
mo e s able blood glucose pla eau anging be ween 90- and 120-min
pos -consump ion, whe eas B-C showed a con inued decline owa d
baseline alues. This p olonged main enance o blood glucose could
esul in se e al me abolic ad an ages. A mo e sus ained glycemic
Fig. 1. In i o cy o oxici y assessmen o B-C and B-5A WSB ex ac s. Viabili y o THP-1-M0 mac ophages ea ed wi h B-C and B-5A WSB ex ac s, a e 24 h (A) and
72 h (B) by he MTT es . Viabili y o undi e en ia ed Caco-2 cells measu ed a e 24 h (C) and 72 h (D) in he same condi ions as (A). Viabili y o en e ocy e-like
Caco-2-de i ed monolaye s measu ed a e 24 h (E) and 72 h (F) in he same condi ions as (A). Resul s a e exp essed as cell pe cen ages ela i e o un ea ed con ol
cells (C) and a e he mean ±SD o wo independen expe imen s in which each condi ion was es ed eigh imes. As e isks indica e signi icance in a pai ed Tukey es
s. C (** p <0.0001). G. Gene exp ession o p o-in lamma o y media o s by qPCR. Gene exp ession analysis o IL-1β, IL-6, IL-8, MCP1 and TNF-
α
in THP-1-M0
mac ophages ea ed o 24 h wi h WSB B-C and B-5A a 200 µg/mL inal concen a ions. The posi i e con ol wi h LPS was a 500 ng/mL inal concen a ion.
Da a a e no malised o he GAPDH and HPRT housekeeping genes. Resul s a e exp essed as mRNA old inc ease compa ed o con ol cells (C) and a e he mean ±SD
o h ee expe imen s pe o med in iplica e. As e isks indica e signi icance in Tukey es s C (ANOVA: IL-1β p <0.05, IL-6 p <0.000001, IL-8 p <0.005, MCP1 p <
0.000001, TNF p <0.0005; Tukey s. C, ** p <0.005).
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
9
in es inal o gano ypic in i o models as well as o mula ion adjus -
men s, such as masking undesi able la o s and op imizing ex u e, o
enhance he pala abili y o A. esculen a-en iched b ead, he eby allow-
ing he inco po a ion o highe amoun s o A. esculen a o achie e
imp o ed bene icial p ope ies o he b ead. Al hough no cy o oxic e -
ec s we e obse ed in b ead con aining Ala ia esculen a, u u e s udies
should quan i a i ely assess he le els o hea y me als and iodine in bo h
he seaweed and inal p oduc . Such analyses would help de e mine
whe he consump ion emains wi hin sa e limi s and ensu es ha long-
e m in ake does no pose heal h isks.
E hical s a emen
The senso y s udy p esen ed in he manusc ip was app o ed by he
E hics Commi ee o he Technical Uni e si y o Cluj-Napoca (Romania,
N . 594/10.04.2024) and adhe ed o he e hical p inciples ou lined in
he 1975 Decla a ion o Helsinki, as e ised in 2013. Be o e he senso y
e alua ion, he pa icipan s we e in o med abou he s udy’s pu pose
and p o ided w i en consen .
We con i m ha his manusc ip has no been published elsewhe e in
whole o in pa and is no unde conside a ion by any o he jou nal.
App o al o all Au ho s’ ins i u ions ha e been g an ed o publish his
wo k. All au ho s ha e app o ed he manusc ip and ag ee wi h i s
submission o Fu u e Foods.
Funding sou ces
This s udy has ecei ed unding om he Eu opean Union’s Ho izon
Eu ope esea ch and inno a ion unding p og amme unde esea ch
p ojec Demons a ion o inno a i e unc ional ood p oduc ion sys ems
based on a mo e sus ainable alue chain o ma ine and eshwa e aw
ma e ials o consciencious Eu opean consume s- NOVAFOODIES, P o-
g amm HORIZON.2.6 - Food, Bioeconomy Na u al Resou ces, Ag icul-
u e and En i onmen , HORIZON.2.6.4 - Seas, Oceans and Inland
Wa e s; Topic: HORIZON-CL6-2022-FARM2FORK-02-05- wo-s age-
Inno a i e ood om ma ine and eshwa e ecosys ems. G an Ag ee-
men No 101084180. doi:10.3030/101084180.
Disclaime
Views and opinions exp essed a e howe e hose o he au ho s only
and do no necessa ily e lec hose o he Eu opean Union o he Eu-
opean Resea ch Execu i e Agency. Nei he he Eu opean Union no
he g an ing au ho i y can be held esponsible o hem.
CRediT au ho ship con ibu ion s a emen
Leona d Mihaly Cozmu a: W i ing – o iginal d a , So wa e,
In es iga ion, Concep ualiza ion. Anca Pe e : In es iga ion. Camelia
Nicula: In es iga ion. Alexand a Ui a asan: In es iga ion. Zsol
Szak´
acs: In es iga ion. Sonia Sca i: Me hodology, In es iga ion.
Maha Moussa: Me hodology, In es iga ion. Se ena Mi a a: Me hod-
ology, In es iga ion. Paula Faja do Be n´
a dez: Me hodology, In es i-
ga ion. Sa a Baldassa i: Me hodology, In es iga ion. Giuliana D a a:
Me hodology, In es iga ion. Anca Mihaly Cozmu a: W i ing – e iew
& edi ing, P ojec adminis a ion, Me hodology, In es iga ion.
Decla a ion o compe ing in e es
The au ho s decla e ha hey ha e no known compe ing inancial
in e es s o pe sonal ela ionships ha could ha e appea ed o in luence
he wo k epo ed in his s udy.
Acknowledgemen s
We would like o exp ess ou since e g a i ude o The Seaweeds
Company, I eland, o p o iding us wi h Ala ia esculen a powde o his
s udy. Thei gene ous con ibu ion has been in aluable, and we g ea ly
app ecia e hei suppo .
Supplemen a y ma e ials
Supplemen a y ma e ial associa ed wi h his a icle can be ound, in
he online e sion, a doi:10.1016/j. u o.2025.100805.
Da a a ailabili y
Da a will be made a ailable on eques .
Re e ences
Ahmad, T., Eapen, M.S., Ishaq, M., Pa k, A.Y., Ka piniec, S.S., S inge , D.N., Sohal, S.S.,
Fi on, J.H., Gu en, N., Ca uso, V., E i, R., 2021. An i-in lamma o y ac i i y o
ucoidan ex ac s in i o. Ma . D ugs 19 (12), 702. h ps://doi.o g/10.3390/
md19120702.
Amigo, J.M., del Olmo, A., Engelsen, M.M., Lundk is , H., Engelsen, S.B., 2021. S aling o
whi e whea b ead c umb and e ec o mal ogenic
α
-amylases. Pa 3: spa ial
e olu ion o b ead s aling wi h ime by nea in a ed hype spec al imaging. Food
Chem. 353, 129478. h ps://doi.o g/10.1016/j. oodchem.2021.129478.
AOAC 945.18-B, 1995. Kjeldahl’s Me hod o P o ein De e mina ion in Ce eals and Feed.
De e mina ion o To al Fa in Flou , B ead, Bake y P oduc s and Pas a wi h
Fig. 4. Va ia ion o P incipal Componen 2 (PC2) as a unc ion o P incipal Componen 1 (PC1).
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
16
P elimina y Acid Hyd olysis. De e mina ion o Ash in Animal Feed. UDK 127
Ope a ing Manual.
AOAC 920.39.C, 1995. The Soxhle Me hod o Ce eal Fa .
AOAC 936.07, 2013. Ash in Flou .
AOAC 925.10, 1925. Solids (To al) and Loss on D ying (Mois u e) in Flou . Ai O en
Me hod.
B´
a cenas, M.E., Rosell, C.M., 2005. E ec o HPMC addi ion on he mic o-s uc u e,
quali y and aging o whea b ead. Food Hyd ocoll. 19, 1037e1043. h ps://doi.o g/
10.1016/j. oodhyd.2005.01.005.
Ba ek, L., Sj¨
olund, A., B ancoli, P., Cica iello, C., Mesi an a, N., N¨
a ¨
anen, E.,
Sche hau e , S., S id, I., E iksson, M., 2025. The powe o p e en ion and
alo isa ion –En i onmen al impac s o educing su plus and was e o bake y
p oduc s a e ail. Sus ain. P od. Consump. 55, 51–62. h ps://doi.o g/10.1016/j.
spc.2025.01.013.
Becke , A., Hill, S.E., Mi chell, J.R., 2001. Rele ance o amylose-lipid complexes o he
beha iou o he mally p ocessed s a ches. S a ch/S a ke 53, 21–30. h ps://doi.
o g/10.1002/1521-379X(200104)53:3/43.0.CO;2-Q.
Begum, R., Howlade , S., Mamun-O -Rashid, A.N.M., Ra iquzzaman, S.M., Ash a , G.M.,
Albad ani, G.M., Sayed, A.A., Peluso, I., Abdel-Daim, M.M., Uddin, M.S., 2021.
An ioxidan and signal-modula ing e ec s o b own seaweed-de i ed compounds
agains oxida i e s ess-associa ed pa hology. Oxid. Med. Cell. Longe . 10, 9974890.
h ps://doi.o g/10.1155/2021/9974890.
Benne , J.P., Robinson, L.F., Gomez, L.D., 2023. Valo isa ion s a egies o b own
seaweed biomass p oduc ion in a Eu opean con ex . Algal Res. 75, 103248. h ps://
doi.o g/10.1016/j.algal.2023.103248.
Cha e , P.I., Wilcox, M.D., Hough on, D., Pea son, J.P., 2015. The ole o seaweed
bioac i es in con ol o diges ion: implica ions o obesi y ea men s. Food Func . 6
(11), 3420–3427. h ps://doi.o g/10.1039/C5FO00293A.
Che y, P., O’Ha a, C., Magee, P.J., McSo ley, E.M., Allsopp, P.J., 2019. Risks and
bene i s o consuming edible seaweeds. Nu . Re . 77 (5), 307–329. h ps://doi.o g/
10.1093/nu i /nuy066.
Commission Regula ion (EC) 152, 2009. Laying Down he Me hods o Sampling and
Analysis o he O icial Con ol o Feed.
Copeland, L., Blazek, J., Salman, H., Tang, M.C., 2009. Fo m and unc ionali y o s a ch.
Food Hyd ocoll. 23, 1527–1534. h ps://doi.o g/10.1016/j. oodhyd.2008.09.016.
Di Giuseppe, D., Sca ì, S., Alessand ini, A., Bassi, A.M., Mi a a, S., Almon i, V.,
Ragazzini, G., Mescola, A., Fila e o, M., A allone, R., Vi ale, G., Scognamiglio, V.,
Gual ie i, A.F., 2022. Acu e cy o oxici y o mine al ib es obse ed by ime-lapse
ideo mic oscopy. Toxicology 466, 153081. h ps://doi.o g/10.1016/j.
ox.2021.153081.
EFSA, 2017. Die a y Re e ence Values o Nu ien s Summa y Repo . Eu opean Food
Sa e y Au ho i y, 2017, e15121. A ailable a 10.2903/sp.e sa.2017. Re ie ed a
25.04.2025 h ps://www.e sa.eu opa.eu/si es/de aul / iles/2017_09_DRVs_summa
y_ epo .pd .
Ena u, B., D ețcanu, G., Pop, T.D., S ǎnilǎ, A., Diaconeasa, Z., 2021. An hocyanins:
ac o s a ec ing hei s abili y and deg ada ion. An ioxidan s 10 (12), 1967. h ps://
doi.o g/10.3390/an iox10121967.
Fanelli, C.G., Lucidi, P., Bolli, G.B., Po cella i, F., 2018. Hypoglycemia. In: Bono a, E.,
DeF onzo, R. (eds) Diabe es Complica ions, Como bidi ies and Rela ed Diso de s.
Endoc inology. Sp inge , Cham. h ps://doi.o g/10.1007/978-3-319-44433-8_22.
Fe uzza, S., Rossi, C., Sca ino, M.L., Sambuy, Y., 2012. A p o ocol o di e en ia ion o
human in es inal Caco-2 cells in asymme ic se um-con aining medium. Toxicol. In
Vi o 26 (8), 1252–1255. h ps://doi.o g/10.1016/j. i .2012.01.008.
Gio dani, M., Mi a a, S., Sca i, S., Passalacqua, M., Fo nasini, L., D a a, G., Meli, M.A.,
Roselli, C., Ma ioli, M., 2025. The cy o oxic/geno oxic ole o impu i ies in soluble
mine als: he case o na u al ( ib ous epsomi e) e sus syn he ic (Epsom sal )
magnesium sulpha e. Sci. To al En i on. 966, 178666. h ps://doi.o g/10.1016/j.
sci o en .2025.178666.
Gua da, A., Rosell, C.M., Benedi o, C., Galo o, M.J., 2004. Di e en hyd ocolloids as
b ead imp o e s and an is aling agen s. Food Hyd ocoll. 8, 241e247. h ps://doi.
o g/10.1016/S0268-005X(03)00080-8.
Hemp , C., Kaise , J.P., Scholde , O., Bue ki-Thu nhe , T., Ho mann, H., Rippl, A.,
Schus e , T.B., Wick, P., Hi sch, C., 2020. The impac o syn he ic amo phous silica
(E 551) on di e en ia ed Caco-2 cells, a model o he human in es inal epi helium.
Toxicol. In Vi o 67, 104903. h ps://doi.o g/10.1016/j. i .2020.104903.
Hlebowicz, J., 2009. Pos p andial blood glucose esponse in ela ion o gas ic emp ying
and sa ie y in heal hy subjec s. Appe i e 53 (2), 249–252. h ps://doi.o g/10.1016/j.
appe .2009.06.014.
Hyns o a, V., S e bo a, D., Klejdus, B., Hedba ny, J., Huska, D., Voj ech, A., 2018.
Sepa a ion, iden i ica ion and quan i ica ion o ca o enoids and chlo ophylls in
die a y supplemen s con aining chlo ella ulga is and spi ulina pla ensis using high
pe o mance hin laye ch oma og aphy. J. Pha m. Biomed. Anal. 148, 108–118.
h ps://doi.o g/10.1016/j.jpba.2017.09.018.
Huwai , E., Al-Saedi, D.A., Mi za, Z., 2022. An i-in lamma o y po en ial o ucoidan o
a he oscle osis: in silico and in i o s udies in THP-1 cells. Molecules 27 (10), 3197.
h ps://doi.o g/10.3390/molecules27103197.
Jim´
enez, A., Fab a, M.J., Talens, P., Chi al , A., 2012. E ec o sodium caseina e on
p ope ies and ageing beha iou o co n. S a ch based ilms. Food Hyd ocoll. 29 (2),
265–271. h ps://doi.o g/10.1016/j. oodhyd.2012.03.014.
Joshi, M., Adhika i, B., Ald ed, P., Panozzo, J.F., Kasapis, S., 2011. Physicochemical and
unc ional p ope ies o len il p o ein isola es p epa ed by di e en d ying me hods.
Food Chem. 129, 1513–1522. h ps://doi.o g/10.1016/j. oodchem.2011.05.131.
J¨
onsson, M., Maube , E., Me kel, A., F ed iksson, C., Ka lsson, E.N., Wendin, K., 2024.
A sense o seaweed: consume liking o b ead and sp eads wi h he addi ion o ou
di e en species o no he n Eu opean seaweeds. A pilo s udy among Swedish
consume s. Fu u e Foods 9, 100292. h ps://doi.o g/10.1016/j. u o.2023.100292.
Kılınç, B., Ci ik, S., Tu an, G., Tekogul, H., Ko u, E., 2013. Seaweeds o ood and
indus ial applica ions. Food indus y. Edi ed by Innocenzo Muzzalupo 10.5772/
53172.
K üge , M.A., Fe e o, C., Za i zky, N.E., 2003. Modelling co n s a ch swelling in ba ch
sys ems: e ec o suc ose and hyd ocolloids. J. Food Eng. 58, 125e133. h ps://doi.
o g/10.1016/S0260-8774(02)00337-0.
Lambadia i, V., Ko akas, E., Tsimihodimos, V., 2020. The impac o die a y glycemic
index and glycemic load on pos p andial lipid kine ics, dyslipidemia and
ca dio ascula isk. Nu ien s 12 (8), 2204. h ps://doi.o g/10.3390/nu12082204.
Li, P., Wang, H., Shao, Q., Kong, B., Qu, X., 2017. Fucoidan modula es cy okine
p oduc ion and mig a ion o THP1de i ed mac ophages ia colonys imula ing
ac o 1. Mol. Med. Rep. 15, 2325–2332. h ps://doi.o g/10.3892/mm .2017.6228.
Lich en hale , H.K., Buschmann, C., 2001. Chlo ophylls and ca o enoids: measu emen
and cha ac e iza ion by UV-VIS spec oscopy. Cu . P o oc. Food Anal. Chem. 1,
F4.3.1–F4.3.8. h ps://doi.o g/10.1002/0471142913. a 0403s01.
Liu, S., Sun, H., Nagassa, M., He, X., Pei, H., Gao, L., Li, X., He, S., 2024. Enhancing b ead
an i-s aling wi h glucose-de i ed Mailla d eac ion p oduc s: in-dep h analysis o
s a ches, glu en ne wo ks, and mois u e s a us. Food Chem. 455, 139760. h ps://
doi.o g/10.1016/j. oodchem.2024.139760.
Ly ou, A.E., Schoina, E., Liu, Y., Michalek, K., S anley, M.S., Panagou, E.Z., Nychas, G.E.,
2021. Quali y and sa e y assessmen o edible seaweeds ala ia esculen a and
saccha ina la issima cul i a ed in Sco land. Foods 10 (9), 2210. h ps://doi.o g/
10.3390/ oods10092210.
MacA ain, P., Gill, C., B ooks, M., Campbell, R., 2007. Nu i ional alue o edible
seaweeds. Nu . Re . 65 (12), 535–543. h ps://doi.o g/10.1301/n .2007.dec.535-
543.
Ma os, J., Ca doso, C., Se alhei o, M.L., Banda a, N.M., A onso, C., 2024. Seaweed
bioac i es po en ial as nu aceu icals and unc ional ing edien s: a e iew. J. Food
Compos. Anal. 133, 106453. h ps://doi.o g/10.1016/j.j ca.2024.106453.
Mihaly Cozmu a, L., Pe e , A., Nicula, C., Jas zębska, A., Jakubczak, M., Pu bayan o, M.
A.K., Bunea, A., Bo a, F., Ui a asan, A., Szak´
acs, Z., Mihaly Cozmu a, A., 2025.
Explo ing he impac o mic owa es, ul asounds, UVA ligh , and X- ays on
polyphenols in algina e–whey p o ein ma ix–encapsula ed ed g ape seed ex ac
powde . Appl. Food Res. 5 (1), 100658. h ps://doi.o g/10.1016/j.
a es.2024.100658.
Mu ugan, A.C., Ka im, M.R., Yuso , M.B.M., Tan, S.H., As as, M.F.B.F., Rashid, S.S.,
2015. New insigh s in o seaweed polyphenols on glucose homeos asis. Pha m. Biol.
53 (8), 1087–1097. h ps://doi.o g/10.3109/13880209.2014.959615.
Na oli, M., Leoni, B.D., D’Agnano, I., Zucco, F., Felsani, A., 2012. Good Caco-2 cell
cul u e p ac ices. Toxicol. In Vi o 26 (8), 1243–1246. h ps://doi.o g/10.1016/j.
i .2012.03.009.
O’Sulli an, A.M., O’Callaghan, Y.C., O’G ady, M.N., Queguineu , B., Hanni y, D.,
T oy, D.J., Ke y, J.P., O’B ien, N.M., 2011. In i o and cellula an ioxidan
ac i i ies o seaweed ex ac s p epa ed om i e b own seaweeds ha es ed in sp ing
om he wes coas o I eland. Food Chem. 126, 1064–1070. h ps://doi.o g/
10.1016/j. oodchem.2010.11.127.
O e duin, J., Jansen, A., 1997. Condi ioned insulin and blood suga esponses in humans
in ela ion o binge ea ing. Physiol. Beha . 61 (4), 569–575. h ps://doi.o g/
10.1016/S0031-9384(96)00504-5.
Phuangbubpha, P., Tha a, S., S iboonaied, P., Sae an, P., Tumnoi, W., Cha oenpanich, A.,
2023. Op imizing THP-1 mac ophage cul u e o an immune- esponsi e human
in es inal model. Cells 12 (10), 1427. h ps://doi.o g/10.3390/cells12101427.
Pole , M., Laloux, L., Cambie , S., Ziebel, J., Gu leb, A.C., Schneide , Y.J., 2020. Soluble
sil e ions om sil e nanopa icles induce a pola ised sec e ion o in e leukin-8 in
di e en ia ed Caco-2 cells. Toxicol. Le . 325, 14–24. h ps://doi.o g/10.1016/j.
oxle .2020.02.004.
Pozzolini, M., Ve gani, L., Ragazzoni, M., Delpiano, L., G asselli, E., Voci, A., Gio ine, M.,
Sca ì, S., 2016. Di e en eac i i y o p ima y ib oblas s and endo helial cells
owa ds c ys alline silica: a su ace adical ma e . Toxicology 361-362, 12–23.
h ps://doi.o g/10.1016/j. ox.2016.07.001.
Pu seys, J.A., Lambe s, L., Delcou , J.A., 2010. Amylose-inclusion complexes: o ma ion,
iden i y and physico-chemical p ope ies. J. Ce eal Sci. 51, 238–247. h ps://doi.
o g/10.1016/j.jcs.2010.01.011.
Qui al, V., Sepúl eda, M., Game o-Vega, G., Jim´
enez, P., 2022. Seaweeds in bake y and
a inaceous oods: a mini- e iew. In . J. Gas on. Food Sci. 28, 100403. h ps://doi.
o g/10.1016/j.ijg s.2021.100403.
Ribo a, P.D., Ausa , S.F., Bel amo, D.M., Leon, A.E., 2005. In e ac ions o hyd ocolloids
and sonica ed-glu en p o eins. Food Hyd ocoll. 19, 93e99. h ps://doi.o g/10.1016/
j. oodhyd.2004.04.018.
Roohinejad, S., Kouba, M., Ba ba, F.J., Saljoughian, S., Amid, M., G eine , R., 2017.
Applica ion o seaweeds o de elop new ood p oduc s wi h enhanced shel -li e,
quali y and heal h- ela ed bene icial p ope ies. Food Res. In . 99 (3), 1066–1083.
h ps://doi.o g/10.1016/j. ood es.2016.08.016.
San os, E., Rosell, C.M., Colla , C., 2008. Gela iniza ion and e og ada ion kine ics o
high- ibe whea lou blends: a calo ime ic app oach. Ce eal Chem. 85, 455e463.
h ps://doi.o g/10.1094/CCHEM-85-4-0455.
Schneide , C.A., Rasband, W.S., Licei i, K.W., 2012. ImageJ (Ve sion 1.47) [Compu e
So wa e]. U.S. Na ional Ins i u es o Heal h. h ps://imagej.nih.go /ij/.
Schwa z, S., Lo enzo, T., 1991. Chlo ophyll s abili y du ing con inuous asep ic
p ocessing and s o age. J. Food Sci. 56, 1059–1062. h ps://doi.o g/10.1111/
j.1365-2621.1991. b14641.x.
Sco , J., Awika, M., 2023. E ec o p o ein–s a ch in e ac ions on s a ch e og ada ion
and implica ions o ood p oduc quali y. Comp . Re . Food Sci. Food Sa . 22,
2081–2111. h ps://doi.o g/10.1111/1541-4337.13141.
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
17
Senadhee a, T.R.L., Hossain, A., Shahidi, F., 2023. Ma ine bioac i es and hei
applica ion in he ood indus y: a e iew. Appl. Sci. 13, 12088. h ps://doi.o g/
10.3390/app132112088.
Si abalan, S., Ross, C.F., Tang, J., Sablani, S.S., 2024. Physical, he mal, and s o age
s abili y o mul ilaye ed emulsion loaded wi h β-ca o ene. Food Inno . Ad . 3 (3),
244–255. h ps://doi.o g/10.48130/ ia-0024-0022.
Song, Y.J., Liu, H.X., Yang, X.G., 2024. The deposi ion o lan hanum ca bona e may
ac i a e mac ophages o induce gas oin es inal mucosal inju y in pa ien s wi h
ch onic kidney disease: an in i o caco-2/THP-1 mac ophage cocul u e model
s udy. J. Biol. Ino g. Chem. 29 (1), 101–112. h ps://doi.o g/10.1007/s00775-023-
02033-x.
Susewind, J., de Souza, C.-W.C., Repnik, U., Collno , E.M., Schneide -Daum, N.,
G i i hs, G.W., Leh , C.M., 2016. A 3D co-cul u e o h ee human cell lines o model
he in lamed in es inal mucosa o sa e y es ing o nanoma e ials. Nano oxicology
10 (1), 53–62. h ps://doi.o g/10.3109/17435390.2015.1008065.
Wang, S., Li, C., Copeland, L., Niu, Q., Wang, S., 2015. S a ch e og ada ion: a
comp ehensi e e iew. Comp . Re . Food Sci. Food Sa e. 14, 568–585. h ps://doi.
o g/10.1111/1541-4337.12143.
Wang, H., Yang, Q., Gao, L., Gong, X., Qu, Y., Feng, B., 2020. Func ional and
physicochemical p ope ies o lou s and s a ches om di e en ube c ops. In . J.
Biol. Mac omol. 148, 324–332. h ps://doi.o g/10.1016/j.ijbiomac.2020.01.146.
Weemaes, C.A., Ooms, V., Van Loey, A.M., Hend ickx, M.E., 1999. Kine ics o chlo ophyll
deg ada ion and colo loss in hea ed b occoli juice. J. Ag ic. Food Chem. 47,
2404–2409. h ps://doi.o g/10.1021/j 980663o.
Wilcox, M.D., Che y, P., Cha e , P.I., Yang, X., Zulali, M., Okello, E.J., Seal, C.J.,
Pea son, J.P., 2021. The e ec o seaweed en iched b ead on ca bohyd a e diges ion
and he elease o glucose om ood. J. Func. Foods 87, 104747. h ps://doi.o g/
10.1016/j.j .2021.104747.
Williams, P.G., 2012. E alua ion o he e idence be ween consump ion o e ined g ains
and heal h ou comes. Nu . Re . 70 (2), 80–99. h ps://doi.o g/10.1111/j.1753-
4887.2011.00452.x.
Willho , E., 2007. Mechanism and heo y o s aling o b ead and baked goods, and
associa ed changes in ex u al p ope ies. J. Tex u e S ud. 4, 292–322. h ps://doi.
o g/10.1111/j.1745-4603.1973. b00844.x.
Xie, L.H., Tang, S.Q., Chen, N., Luo, J., Jiao, G.A., Shao, G.N., Wei, X.J., Hu, P.S., 2014.
Op imisa ion o nea -in a ed e lec ance model in measu ing p o ein and amylose
con en o ice lou . Food Chem. 142, 92–100. h ps://doi.o g/10.1016/j.
oodchem.2013.07.030.
Zeb, A., 2012. Oxida ion and o ma ion o oxida ion p oduc s o β-ca o ene a boiling
empe a u e. Chem. Phys. Lipids 165 (3), 277–281. h ps://doi.o g/10.1016/j.
chemphyslip.2012.02.005.
Zhao, Q., Lin, J., Wang, C., Yousa , L., Xue, Y., Shen, Q., 2021. P o ein s uc u al
p ope ies and p o eomic analysis o ice du ing s o age a di e en empe a u es.
Food Chem. 361, 130028. h ps://doi.o g/10.1016/j. oodchem.2021.130028.
L. Mihaly Cozmu a e al.
Fu u e Foods 12 (2025) 100805
18
