Journal Pre-proof

Bioactive-Loaded Nanocarriers for Functional Foods: From Designing to

Bioavailability

Cristian Dima, Elham Assadpour, Stefan Dima, Seid Mahdi Jafari

PII: S2214-7993(19)30090-6
DOI: https://doi.org/10.1016/j.cofs.2019.11.006
Reference: COFS 522

To appear in: Current Opinion in Food Science

Please cite this article as: Dima C, Assadpour E, Dima S, Jafari SM, Bioactive-Loaded
Nanocarriers for Functional Foods: From Designing to Bioavailability, Current Opinion in Food
Science (2019), doi: https://doi.org/10.1016/j.cofs.2019.11.006

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 Bioactive-Loaded Nanocarriers for Functional Foods: From Designing to
Bioavailability
Cristian Dimaa, Elham Assadpourb, Stefan Dimac, Seid Mahdi Jafarib*

a
“Dunarea de Jos” University of Galati, Faculty of Food Science and Engineering, Domneasca Street, 111,
RO-800201, Galati, Romania
b
Department of Food Materials and Process Design Engineering, Gorgan University of Agricultural Sciences
and Natural Resources, Gorgan, Iran
c
“Dunarea de Jos” University of Galati, Faculty of Science and Environment, Domneasca Street, 111, RO-
800201, Galati, Romania
*Corresponding author: Seid Mahdi Jafari (smjafari@gau.ac.ir)

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Abstract

In recent years, researchers' concern for the applications of nanotechnologies in the food industry has grown

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exponentially. This has led to a major change in both consumer and producer behaviours. Thus, recent findings
have shown that consumers increasingly prefer value-added foods, which provide significant health benefits.
In this context, new approaches have emerged in the design of functional foods, such as the “reverse
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engineering” approach that ensures the necessary conditions for designing sustainable "nano-food" systems,
in accordance with the requirements of the consumer. The stages of nano-food design using the reverse
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engineering method, the factors affecting the bioavailability of nanoencapsulated biocomponents and the
fortification of foods with bioactive-loaded nanocarriers are briefly analysed in this review.
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Keywords: food-nanocarriers; reverse engineering; bioavailability; design; functional foods.

1. Introduction
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The development of nanotechnologies has allowed the implementation of new advances in agrifood sectors,
such as: designing “green” nanomaterials (nanopesticides, nanofertilizers), contributing to the increase of crops
(cereals), fruits and vegetables, or feed for livestock; manufacturing innovative nanodelivery systems of
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biocomponents able to contribute to a better food quality and safety, as well as bioavailability of active
compounds; producing intelligent packaging based on nanomaterials able to provide high protection for foods;
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and inventing high-performance nanosensors, able to detect traces of contaminants, mycotoxins and
microorganisms in foods [1●●].

Nowadays foods are not just the sustenance for human life, but have also become the main ways to prevent
and treat chronic illnesses. That is why designing such foods is a complex process in which the consumer takes
part alongside doctors, technologists, biologists, economists, sociologists, etc. [2●●]. Applying nanomaterials
in the food product sector is in agreement with the general principles of durable development, specific to the
globalisation phenomenon [3●- 5]. The concept of sustainability integrates foods and food systems into a chain
of seventeen sustainable development goals formulated by UNO, whose main purpose is to provide decent
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living conditions and equitable access to food for all people on Earth, who should live in peace, harmony, and
respect towards the environment [6]. In this respect, de Vries et al. [7●●] identified three main directions of
development for food products:
- increasing the efficiency of food processing technologies;
- developing innovative new products based on bioactive-loaded nanocarriers which can contribute to
superior bioavailability and disease prevention and treatment;
- identifying socioeconomic and cultural factors involved in food preparation processes.

Thus, there have been advancements in research on rational exploitation of ecosystems and developing new
food-nanomaterials which are less harmful to the environment; reducing the consumption of water and energy,
and last but not least, reducing the amount of discarded food. It is estimated that a third of the annual food
production (~1.3 billion tonnes) gets lost or wasted [8]. This wasted food, which may improve the daily diet
of many Earth inhabitants, become dangerous waste to the environment. This review highlights the status of

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nanomaterials in the context of requirements to develop new sustainable foods, characterized by high
nutritional and health performance. It also provides a general presentation of all the stages involved in the

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reverse engineering design of bioactive-loaded nanocarriers in functional foods, the factors influencing the
bioavailability of nanoencapsulated biocomponents, and their applications in food functionalization.
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2. Reverse engineering in the design of functional food products with nanocarriers

Today, due to demands for the sustainable development of societies (environment, health and economy), food
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design has been changed from classic design (based on forward engineering approach, when foods were
manufactured according to technology and raw materials), placing the consumer at the end of food chain, to a
modern design (based on reverse engineering, where the initial stage is the consumer’s preferences) and the
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“anamnesis” of the foods required by the consumer [7●●]. The use of Multi-Criteria Reverse Engineering as a
design method for functional foods based on encapsulates is a complex process, centred on the consumer [9●●].
This design method takes into account the following aspects: performing a nutritional, sensorial, and functional
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evaluation of the food product according to the consumer’s requirements; selecting the target bioactives,
nanocarrier materials and nutraceuticals delivery systems, encapsulation technique, food matrix or the
excipient; choosing the packaging method; analysing the costs; the impacts on environment; analysing the
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perception of food product; and finally, testing the bioavailability of nutraceuticals incorporated within food.
Consumers are aware of the role of functional foods, foods for particular nutritional uses (PARNUTs) and
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medical foods in preventing and treating certain illnesses, like obesity, type II diabetes, cardiovascular
diseases, immunodeficiency, cancers, etc. [10●●].
Fig. 1 shows the main stages involved in the traditional design and in reverse engineering design of functional
foods.
Fig. 1

In recent decades a high number of researchers have focused on developing new nanocarriers with possible
applications in manufacturing functional foods. Unfortunately, a few products have been commercialized in
this regard. For example, out of the numerous nanomaterials studied, only fifty-five have been accepted by
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European Food Safety Authority (EFSA, 2014: EN-621)) for use in the agrifood sectors including: inorganic
nanomaterials (silver, iron, calcium, magnesium, selenium, titanium oxide, etc.) and organic nanomaterials
(lipid-, protein- and polysaccharide-based nanomaterials) [1●●, 3●●]. This is due, on the one hand, to the
international non harmonisation of regulations in food nanotechnology, and on the other hand, to the
insufficient and inconclusive studies on the toxicity of nanomaterials in foods [1●●, 2●●]. Most studies on the
toxicity of food nanomaterials are performed in vitro, testing nanomaterials that were free or introduced in
model food systems, and far fewer tests are performed in vivo with nanomaterials in real food matrices [11●●].
Similarly, it was found that the consumer is cautious and sceptical about nanomaterial-based functional foods
[12].
2.1. Selection of the biocomponents
The main criterion in choosing target bioactive components is biological activity, which should correspond to
the needs of final consumer. In this respect, our research teams have studied the extraction, biological activity

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and encapsulation of a large variety of bioactive compounds, such as carotenoids, polyphenols, vitamins,
minerals, phytosterols, polyunsaturated fatty acids (PUFAs), essential oils, etc., that may contribute to
preventing or treating chronic diseases [13●●]. Also, encapsulation of biocomponents takes into account their

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physicochemical characteristics, such as: the aggregation state, crystallinity, molecular weight, chemical
structure, solubility, lipophylicity, the repartition constant (P and log P), etc. These characteristics mainly
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impact the bioaccessibility and bioavailability of bioactive compounds. Briefly, the famous “rules of five”,
state that a bioactive product has a good oral bioavailability if it meets three of the following four requirements:
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M ≤ 500g/mol; logP ≤ 5; hydrogen bond donors: HBD ≤ 5; hydrogen bond acceptors: HBA ≤ 10 [14].
2.2. Selection of the nanocarrier materials
The selection of encapsulating materials influences the characteristics of nanoparticles (encapsulation
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efficiency, size and particles size distribution, surface morphology and charge, release mechanism),
nutraceutical bioaccessibility and bioavailability. To encapsulate nutraceuticals, only food-grade materials
accepted as GRAS (Generally Recognized As Safe) are used, such as proteins, lipids, and polysaccharides
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[11●●].
The other selection criteria of nanocarrier materials refer to: i) physicochemical and biological characteristics
(colour, taste, odour, resistance to pH, temperature, oxygen, light, moisture variation, low chemical reactivity,
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minimum toxicity) that should not affect the food sensorial attributes; ii) functional properties (solubility,
viscosity, surface tension, electric charge, gelling ability, mechanical resistance); iii) sources (natural,
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synthetic, semisynthetic); iv) accessible costs; v) cultural and nutritional restrictions (religious restrictions;
vegetarian and raw vegan diets) [15].
Similarly, the selection of food-nanocarrier materials is also performed in accordance with the food matrix in
which bioactive-loaded nanoparticles are introduced. Thus, lipid-based nanoparticles such as nanoemulsions,
solid lipid nanoparticles (SLNs), and nanostructured lipid nanocarriers (NLCs) are excellent delivery systems
for lipophilic bioactives (carotenoids, antioxidants, vitamins, antimicrobials, flavours, colours, etc.) used in
beverages fortification, due to the optical properties (optically transparent nanoemulsions), physical stability,
and high oral bioavailability.
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The most common proteins used as encapsulating materials are milk proteins (casein, caseinates, whey
proteins, β-lactoglobulin, lactoferrin, etc.), and vegetable proteins (soy proteins, zein, gliadin, etc.). Protein-
based nanocarriers are generally spherical, but they may also be shaped differently, such as fibrillary or
nanotube. Protein-based nanocarriers may encapsulate both lipophilic and hydrophilic biocomponents. The
advantage of proteins consists in their polyelectrolyte structure providing nanoparticles with a different electric
charge, according to pH. Thus, for pH values under the isoelectric point (pI), proteins have a net positive
charge, while for higher pH values, the protein charge becomes negative. This behaviour of proteins influences
the bioavailability of biocomponents and allows the preparation of nano-laminated systems via layer-by-layer
technique [16].
Polysaccharides are natural biopolymers extracted from vegetable sources (starch, cellulose, alginates,
carrageenans, etc.), animal sources (chitosan), or derived by microbiological processes (xanthan). Some
polysaccharides are neutral polymers (starch, cellulose), and others have a polyanion structure (alginates,

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pectins, carrageenans). Due to the negative charge, polysaccharides may interact with proteins, at values under
pI forming electrostatic complexes, or may form gels in the presence of some metallic ions (Ca2+, Zn2+, K+,

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etc.) [17, 18]. Similarly, certain polysaccharides are digestible (starch), while others are not (alginate, pectin).
Nanoparticles prepared from digestible polysaccharides are broken down under the action of amylases in the
mouth, stomach, or small intestine, allowing the release of encapsulated biocomponents in these GIT segments;
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while nanoparticles prepared from non-digestible polysaccharides are resistant to these enzymes, but are
degraded in the colon by fermentation in the presence of microbiota.
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2.3. Selection of the encapsulation technique/nanocarrier type
Food-nanocarriers may be prepared in two manners: by fragmenting materials up to the nanometric scale (top
down methods: e.g. grinding of solid materials, preparation of nanoemulsion by high-energy methods, etc.) or
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by associating molecules in nanoaggregates (bottom up methods: e.g. self-assembling of surfactants in

micelles, vesicles, liposomes, etc.). The selection of encapsulation methods is performed according to the type
and characteristics of the nanocarriers that should be compatible with the food matrix in which they are to be
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introduced. Thus, some techniques are used for the preparation of liquid nanocarriers, such as nanoemulsions
used in drinks or meat products, while other techniques are used for the preparation of solid nanocarriers, such
as powders of polymeric nanocarriers used in the fortification of dairy products, cereals and baked goods. The
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type of biocomponents strongly influences the choice of encapsulation technique. Thus, hydrophobic
components are encapsulated by techniques specific to the preparation of lipid-based nanocarriers, while
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hydrophilic components are encapsulated in other colloidal delivery systems, such as: molecular complexes
(protein-polyphenol complexes, polyphenol-cyclodextrin complexes), water-in-oil-in-water double emulsions,
multilayer emulsions, liposomes, biopolymeric nanocarriers, etc. Other important factors taken into account
when selecting the encapsulating techniques of biocomponents are as follows: the complexity degree of the
technology, the necessary time to manufacture of nanocarriers, cost, and environmental impact.
3. Nanocarrier characteristics and bioavailability of loaded bioactives
Numerous papers on bioactive-loaded nanocarriers have focused on their behaviour in the gastrointestinal tract
(GIT) segments, and also their bioavailability evaluation [24-38] (Table 1). Oral bioavailability is a measure
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of the bioefficiency of bioactive compounds, being the fraction of ingested bioactive amount that becomes
available to manifest its bioactivity at the level of body organs and tissues. Oral bioavailability is the result of
processes that the biocomponents undergo within four GIT segments: mouth, stomach, small intestine and
large intestine (colon).
Table 1
McClements and his team identified three important processes involving bioactive-load nanocarriers which
define oral bioavailability: bioaccessibility, absorption and transformation [20]. Each of these processes
contains factors that limit bioavailability (Fig. 2). Based on the influence of limiting factors of bioavailability,
they proposed a nutraceutical classification scheme known as the Nutraceutical Bioavailability Classification
Scheme or NuBAC [20●●]. The studies carried out in recent years have investigated the factors influencing the
oral bioavailability of bioactives, the handling techniques of nanocarrier characteristics in order to increase the
bioavailability of bioactives, and the development of in vitro and in vivo techniques to assess oral

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bioavailability [21, 22].
This section will deal with the important factors affecting the oral bioavailability of encapsulated

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biocomponents.
Fig. 2

3.1. Composition and loading capacity -p

The chemical composition of nanocarriers influences in a decisive manner the release and solubility of
biocomponents in the GIT segments. For example, the release and solubility of hydrophobic bioactives from
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lipid-based nanocarriers are favoured by the action of lipase in mouth, stomach and intestines, which breaks
down triacylglycerides (TAGs) into free fatty acids (FFAs) and monoacylglycerides (MAGs); thus
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contributing, together with the bile salts, to the solubilisation of bioactives into mixed micelles [18]. Research
has shown that lipid-based nanocarriers have a different digestibility in the GIT, depending on the chemical
composition of lipids. Thus, lipids with a high content of medium chain triglycerides (MCTs) are easier to
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digest than those with a higher content of long chain triglycerides (LCTs); but the bioavailability of vitamin
D3 in LCT-prepared nanoemulsions (corn oil and fish oil) was higher than in MCT-prepared emulsions, as the
solubility of vitamin D3 in free fatty acid micelles derived from LCT was higher than those micelles derived
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from LCTs [22].

Also, the amylases and proteases hydrolyse starch and proteins, determining the release of bioactives from
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polymeric nanocarriers [21]. Other materials, such as resistant starch, pectins, alginates, etc. are resistant to
the action of enzymes and pH variation, ensuring the release of bioactives in the colon, where these materials
are degraded by bacteria [23].
The loading capacity of nanocarriers affects the amount of nanocarriers introduced in the food, and the
necessary dose for the bioactive effect of the biocomponents.
3.2. Size and surface
The size and electric charge of bioactive-loaded nanocarriers are important factors affecting colloidal stability,
release rate and absorption through the intestine wall. So, certain authors investigated the physical changes of

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the nanoemulsions in GIT segments, evincing the presence of flocculation and coalescence. For instance,
Salvia-Trujillo and his colleagues studied the influence of oil droplet size on β-carotene bioaccessibility,
showing that the decrease of oil droplet size led to the increase of bioavailability, due to the increase of
interphase surface that the lipase accessed to hydrolyse TAGs [21, 31]. Also, size and charge affect the uptake
of nanocarriers through tight junctions or active and passive transport mechanisms (Fig. 2). Thus, hydrophobic
surface nanocarriers pass through the lipophilic bilayer of enterocyte cells more easily than hydrophilic surface
nanocarriers and positively-charged nanocarriers pass more easily than negatively charged ones [39].
3.3. Food matrix effects and GIT fate of nanocarriers
In preparing functional foods, bioactive-loaded nanocarriers are dispersed in various food matrices, such as:
beverages, yogurt, cheese, bread, etc. that contain a multitude of chemical compounds: fats, proteins,
carbohydrates, minerals, antioxidants, colouring agents, etc. In the food matrix there are various interactions
between these components that ensure the texture and the sensorial and nutritional characteristics of the food,

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and influence the bioactivity of biocomponents. Thus, the food matrix effect explains why the biological
activity of a functional food on the whole is not equal to the sum of the biological values of its biocomponents,

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or why the biological activity of the same biocomponents is different when ingested in various foods [40]. The
food matrix effect influences the GIT fate of bioactive-loaded nanocarriers. This happens because in GIT some
components, like proteins, mineral ions, and surfactants from the food matrix along with the GIT fluids are
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adsorbed on the surface of nanoparticles, changing the size, charge and hydrophobicity, and affecting the
bioavailability of biocomponents and their transport through the mucus layer and intestinal epithelium cells
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[19●●, 22].

4. Application of bioactive-loaded nanocarriers in the food industry

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In the past few years, more and more researchers studied the effect of bioactive-loaded nanocarriers on the
sensorial, physicochemical and biological characteristics of real food matrices, for the purpose of identifying
their potential applications in the food industry [41].
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4.1. Dairy products

Since milk products are constantly consumed in high amounts, they were used as food matrices for
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enhancement with the biocomponents necessary to keep the population healthy [40]. Researchers have studied
various ways of fortifying dairy products, such as: using natural products (plants, seeds, nuts, fruits, etc.), using
free phytocomponents extracted from vegetables (phytosterols, vitamins, carotenoids, polyphenols, PUFA ω-
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3, etc.) or dispersing certain bioactive-loaded nanocarriers [42]. As an example, symbiotic dairy products have
been prepared by means of prebiotics polysaccharide-based nanocarriers loaded with probiotic bacteria.
Polysaccharide prebiotics are indigestible fructo-oligosaccharides (inulin, oligofructose) which provide
development conditions for probiotic bacteria [43].
4.2. Meat products

Meat and meat products are an important source of nutrients for the consumers’ health. The use of
nanomaterials in meat industry targets the following: reducing the content of components that may affect

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consumer health, such as fats, salt, nitrites; delivery of biocomponents with antioxidant or antibacterial activity;
and manufacturing packaging able to provide increased safety to meat and meat products [44]. In this respect,
certain authors used nanoemulsions and double emulsions as both nanocarriers for the delivery of hydrophobic
bioactives (essential oils, PUFAs), and systems of replacing fat with vegetable oils (olive oil, linseed oil, corn
oil, etc.) [45, 46].
4.3. Beverages

Soft drinks and fruit juices are the most consumed beverages, bringing notable benefits to manufacturers. Soft
drinks contain a large variety of hydrophobic compounds, like flavour oils, nutraceuticals, colorants, oil-
soluble vitamins, antimicrobials, clouding agents and weighting agents. For water dispersion, these compounds
were encapsulated in various types of nanocarriers, such as: nanoemulsions, double emulsions, SLNs, and
liposomes. [47]. Fruit juices are natural drinks with a different content of biocomponents contributing to
consumer health [48●●]. In order to increase the nutritional value, physical and microbiological stability,

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various bioactive-loaded nanocarriers have been added into fruit juices, such as: liposomes loaded with α-
tocopherol, chlorogenic acid, ascorbic acid, nanoemulsions loaded with nisin and thymol, and polymeric

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nanoparticles loaded with trans-cynamaldehyde, enzymes, probiotics etc. [ 48●●].
4.4. Bakery products

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Among bakery products, bread is the most consumed, so many studies have focused on its enhancement with
different biocomponents, like n-3 PUFAs, vitamins, and minerals. Bread enhancement with fish oil as a main
source of n-3 PUFAs required its encapsulation in various types of nanocarriers, due to its unpleasant flavour
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and low stability to light and oxygen. Thus, Ojagh and Hasani [49] prepared fish oil nanoliposomes (~300nm
in size), with the encapsulation efficiency ~ 90%, which they used in bread fortification without modifying its
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texture and sensorial attributes. Other authors reported encapsulation of thyme (Thymus vulgaris) essential as
a natural preservative in bakery product [50].
5. Conclusion
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The durable development of the food sector in general, and of the functional foods in particular, require the
strict observance of three main factors (economy, health, environment), and designing functional foods by the
reverse engineering method which considers the consumer as the main link in food design and preparation.
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Although the market is relatively poor in “nanofoods”, the number of scientific papers on preparing and
characterizing new bioactive-loaded nanocarriers, developing high performance techniques of in vitro and in
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vivo assessment of the oral bioavailability and testing the nanomaterial toxicity risk, has increased
exponentially in recent years. This study provided an overview of different bioactive-loaded nanocarriers and
their bioavailability and application in production of functional foods.

Conflict of interest
All authors declare that there is no conflict of interest.

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Acknowledgement

This work was supported by an internal research grant of “Dunarea de Jos” University of Galati, Romania,
contract number GI -02/01.03.2018.

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enzymes, and probiotics for fruit juice fortification, preservation, and processing: An overview. J Funct
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**An excellent review article on different bioactive-loaded nanocarriers used in fruit juice
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49. Ojagh SM, S Hasani S: Characteristics and oxidative stability of fish oil nano liposomes and its
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Encapsulated thyme (Thymus vulgaris) essential oil used as a natural preservative in bakery product. Food
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Fig. 1. Forward engineering and reverse engineering approaches used in the design of foods and functional foods
containing bioactive-loaded nanocarriers
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Fig. 2. Gastrointestinal fate of functional foods containing bioactive-loaded nanocarriers and the main factors limiting
the oral bioavailability of bioactives: Bioaccessibility (Liberation, Solubilization, Interactions), Absorption (Mucus
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layer, Tight junction transport, Infflux (active) transporters, Efflux transporters), and Transformation (Chemical
degradation, Metabolism).
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 Table 1. Overview of bioaccessibility and bioavailability of bioactive-loaded nanocarriers
Bioactive Nanocarriers Characteristics
compound of nanocarriers
Curcumin Inclusions complexes/ Curcumin solubility in
4-α-glucanotransferase- complexes nanocarriers has
modified rice (GS) increased by approximately
β-cyclodextrin (CD) 2,241- 2,846-fold,
maltodextrin (MD) compared to pure curcumin
Curcumin WPI-nanoemulsions; WPI-nanoemulsions:
WPI/chitosan Size ~ 186nm
multilayers Zp ~ - 52 mV
nanoemulsions WPI/Chitosan multilayers
nanoemulsions:
Size~5700 nm
Zp ~ +40 mV
Epigallocatechin Nanostructured lipid Size ~300nm
gallate (EGCG) carriers Polydispersity < 0.2
Zeta potential ~ - 30 mV
Encapsulation efficiency~
90%
Phytosterols Nanoporous Corn and
Wheat Starch
Impregnation capacity:
100-180 mg phytosterol/g
Bioaerogels gel
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Nanoporous starch Impregnation capacity of
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Phytosterols aerogels (NSA) in the PS-NSA: 91.2 ± 1.9 mg
granola bars and phytosterols/g PS-NSA
puddings
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Droplet size:
β-carotene Nanoemulsions Starch-stabilized
nanoemulsion: 180 nm
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Tween-stabilized
nanoemulsion: 195 nm
Caseinate-stabilized
nanoemulsiilor:205 nm
Lycopene Whey protein isolate Particles size: 107.8-348.2
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nanoparticles nm
Polydispersity: 0.1-0.3
Loading capacity: 8.20-
12.3%
Encapsulation efficiency:
53.7-64.7%
Eicosapentaenoic Nanoemulsions Droplet size: ~89 nm
acid (EPA) and Zeta potential: ~ 12.5 mV
docosahexaenoic Polydispersity: ~ 0.2
acid (DHA)
Chia seed oil Chia seed Particles size:
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Bioaccessibility Ref
(BA)/Bioavailability
Simulated digestion model (stomach, intestine);
Bioaccessibility:
GS-cur: 2.73 ± 0.14% [24]
MD-cur: 1.75 ± 0.32%
CD-cur: 1.55 ± 0.50%
Cur pure: 0.24 ± 0.01%
Dynamic gastrointestinal model (stomach,
duodenum, jejunum and ileum):
WPI-nanoemulsions BA=29.8±0.5%,
WPI/chitosan multilayers nanoemulsions [25]
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BA=37.2±7,6%
Caco-2 cells as “in vitro” intestinal absorption
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model;
Apparent permeability (Papp): [26]
EGCG free: 1.35·10-6 cm s-1;
EGCG-loaded NLC: 1.28 ·10-6 cm s-1;
Folic acid-functionalized NLC:
-p
2.38·10-6cm· s-1.
Dynamic gastrointestinal model (oral, gastric
and intestinal digestion)
BA phytosterols free: 1.2-1.4% [27]
BA phytosterols in monolith gels:
14.3-27.7%
BA phytosterols in powdered gels: 7-10%
Oral, gastric, and intestinal digestion model;
Bioaccessibility:
Crude phytosterols: 28.0 - 29.8 % in low-fat
granola bars and 31.3–31.5% in regular-fat
granola bars; [28]
Low crystallinity phytosterol nanoparticles:
88.2% in low-fat granola bars, 91.8% for
regular-fat granola bars, and 19.2% for
puddings
“In vitro” multistage simulated GIT model
Bioaccessibility:
Starch-stabilized nanoemulsion: 8.10% [29]
Tween-stabilized nanoemulsion: 13.80%
Caseinate-stabilized
nanoemulsion: 29.80%
“In vitro” cellular uptake using
the MCF-7 cells model. [30]
The uptakes after 3h of incubation:
Free lycopene: 24.18%;
Lycopene nanoparticles: 46.7%
“In vivo” experimental model [31]
Absorption in intestinal everted sac model in
rats
Trans epithelial absorption (90min):
Duodenum: ~ 2 mg/cm
Jejunum: ~ 5.5 mg/cm
Ileum:~ 5 mg/cm
“In vitro” static digestion model [32]
 protein-gum complex Initial: 18-20μm
coacervates Gastric condition: 180-
200μm
Zeta potential:
Initial ~ -5 mV to – 10 mV
GIT stages: ~ -10 mV to –
20 mV
Whey protein WPC-Fe nanoparticles was
Fe concentrate (WPC) prepared by spray drying
nanoparticles method;
Yield: 8.21 g/100 mL
Solubility: 87.53%
Iron content: 11.042 mg
Fe/g powder
Vitamin A Sodium caseinate nano- The complexes were freeze
complexes: dried and reconstituted
Succinylated sodium in water for in-vitro
caseinate (SNaCaS); digestion.
Reassembled sodium
Caseinate (RNaCaS);
Reassembled-
succinylated sodium
caseinate (RSNaCaS)
Vitamin D3 Pea protein-stabilized Emulsion droplets size:
nanoemulsions 170-350 nm
Zeta potential ~ - 25 mV
The influence of pressure,
homogenization cycles, oil
and protein concentration
was studied;
Encapsulation efficiency:
94.8-95.3%
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Vitamin D3 Whey protein-stabilized VD3-Nanoemulsions were
nanoemulsions prepared with: digestible oil
(DO), indigestible oil (IO),
DO+IO mixtures and whey
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protein isolate
Vitamin D3 Composite gels Whey protein isolate and
lotus root amylopectin
composite gels
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Vitamin D3 W/O/W double W/O/W double emulsion:
Droplet size: 11-20μm
Zeta potential: -15 mV to -
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25 mV
Polymer nanoparticles with
W/O/W double emulsion:
Particles size: 850-900 μm
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Zeta potential: -15 mV to
+9 mV
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Free fatty acid (FFA) after 1h digestion:
400 – 900 mg FFA/g oil
“In vivo” experimental model
Apparent digestibility coefficient in anaemic [33]
rats group:
62.53% at 7 days and 80.12% at 28 days
Retention/intake in anaemic rats group:
62.49% at 7 days and 80.10 at 28 days
WPC-Fe complex supplementation
decreased LDL cholesterol
“In vitro” semi dynamic simulated digestion
model (Bioaccessibility %)
Free VA: 68%
SNaCaS-VA: 90%
RNaCaS-VA: 87.67% [34]
RSNaCaS-VA: 90.91%
VA uptake by Caco-2 cells (Bioavailability)
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Free VA: 30.29%
SNaCaS-VA: 37.04%
RNaCaS-VA: 39.60%
RSNaCaS-VA: 45.14%
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Cell uptake study (Bioavailability)
Caco-2 cells as “in vitro” intestinal absorption
model [35]
Uptake efficiencies:
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~ 100% after 2h and 62.5 μg/ml nanoemulsion
concentration;
~ 60% after 2h and 31.25 μg/ml nanoemulsion
concentration
Static “in vitro” digestion model [36]
Bioaccessibility:
VD3-Nanoemulsions (DO): ~65%
VD3-nanoemulsions (IO): ~ 20%
VD3-nanoemulsions (DO+IO): ~45%
VD3 release from gels in simulated intestinal
fluid: 8.9% (6h, 37oC)
“In vivo” intestinal absorption of vitamin D3 [37]
(eight-week-old male C57BL/6 mice)
Bioavailability (at 30 days):
VD3 free: 15%
VD3-composite gels: 35%
“In vitro” multiple stage digestion model:
Bioaccessibility of Ca: 40-70%
“In vitro” single stage digestion model:
Bioaccessibility of Ca: 50-80% [38]
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