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In vitro intestinal absorption of carotenoids delivered as molecular inclusion complexes with b-cyclodextrin is not inhibited by high-density lipoproteins – PowerPoint PPT presentation

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1
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes with
b-cyclodextrin is not inhibited by high-density
lipoproteins Elisabet Fernández-García, Irene
Carvajal-Lérida, Francisco Rincón, José J. Ríos
and Antonio Pérez-Gálvez
aperez_at_cica.es Food Biotechnology
Department Instituto de la Grasa (CSIC) Av. Padre
García Tejero 4, 41012 Sevilla (SPAIN)
2
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • SIGNIFICANCE OF BIOAVAILABILITY STUDIES
  • Interest in the screening of bioavailability has
    increased for different reasons
  • Existence of undernourished population
  • Epidemiological studies have associated between
    consumption of fruit and vegetables to a lower
    risk of developing degenerative diseases
  • Development of food products with added
    nutritional value
  • Food legislation concerning functional foods

3
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • A MULTIFACTORIAL SYSTEM EFFECTS CAROTENOID
    ASSIMILATION
  • Carotenoids are fat soluble compounds
  • Liberation from food matrix
  • Incorporation to mixed micelles
  • Absorption by epithelial cells through
    simple/facilitated diffusion mechanisms
  • Absorption efficiency is relatively low from
    fruits and vegetables
  • Fiber, kind and amount of fat, interaction among
    carotenoids
  • Increase of absorption efficiency from processed
    fruits and vegetables (homogenization and thermal
    processing)

4
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • INTER-INDIVIDUAL VARIABILITY AND THE
  • NON-RESPONDER CONCEPT
  • Comparison of the in vivo lutein absorption
    efficiency non-responder versus
    lutein-responders group

Responders group
non-Responders group
5
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • AIM OF THE STUDY
  • Estimation of the bioaccessibility of dietary
    carotenoids reached when they are delivered as
    inclusion complexes
  • Dietary carotenoids (b-carotene, lutein and
    lycopene) were formulated as micellar solutions
    (control) or inclusion complexes with
    b-cyclodextrin
  • BBMVs preparations were used as the in vitro
    model to assay carotenoid uptake from both
    carotenoid formulations (micellar solution or
    carotenoid-CyDIC) at three concentration levels
  • Comparison of absorption efficiency under
    inhibition conditions of membrane protein
    transporters (BBMVs pre-incubated with HDLs)

6
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotenoid absorption
    efficiency in function of the concentration and
    delivering method

Assimilation of b-carotene
  1. Saturation versus linear trend
  2. Increase of efficiency at 2.5 mM

7
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotenoid absorption
    efficiency in function of the concentration and
    delivering method

Assimilation of lycopene
  1. Saturation versus linear trend
  2. Increase of efficiency at 1.0 mM

8
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotenoid absorption
    efficiency in function of the concentration and
    delivering method

Assimilation of lutein
  • Saturation versus linear trend
  • Increase of efficiency at 2.5 mM
  • A lower absorption efficiency
  • was observed versus carotenes

9
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Primary effects of the factors concentration,
    donor solution type and inhibition

10
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • SUMMARY
  • Factors concentration and donor solution type
  • Association between concentration and
    assimilation mechanism
  • Structural features (polarity) or different
    affinity of transporters may explain the
    absorption efficiency data of carotenes and
    lutein
  • Significant increase on efficiency of the
    assimilation is reached when carotenoids were
    delivered as inclusion complex with CyD

11
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Does delivery of carotenoids as inclusion complex
    mean an increase on absorption efficiency?
  • Absorption rate in pmol/(mg protein x min)

12
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • SUMMARY
  • Factors concentration and donor solution type
  • At the lowest concentration the carotenoids from
    micellar solutions were more efficiently
    assimilated
  • At 1.0 mM a heterogeneous behavior was observed
  • Only at the highest concentration, carotenoids
    from inclusion complex solutions were more
    efficiently assimilated in comparison with the
    carotenoid micellar solutions at that
    concentration (b-Car 51 Lut 185 Lyc 128).
    What absorption mechanism does apply for
    inclusion complexes?

13
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Two-stage mechanism for carotenoid assimilation
    from inclusion complex solutions release and
    absorption

Lysate of BBMVs after assimilation procedure with
lutein inclusion complex at the donor solution
Lutein inclusion complex at the donor solution
14
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Solubility of carotenoids is a rate-limiting step
    of absorption.
  • Dissolution kinetics of the complex is enhanced
    at high concentrations and depends on binding
    constant of the host-guest complex


K complexation
De-complexation
Assimilation Passive or facilitated diffusion
15
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotenoid absorption
    efficiency in function of the concentration and
    delivering method with inhibitor

Assimilation of b-carotene
  1. Decrease of 50 (mean value)
  2. Saturation versus linear trend
  3. Increase of efficiency at 0.5 mM

16
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotenoid absorption
    efficiency in function of the concentration and
    delivering method with inhibitor

Assimilation of lycopene
  1. Decrease of 40 (mean value)
  2. Saturation versus linear trend
  3. Increase of efficiency at 0.5 mM

17
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotenoid absorption
    efficiency in function of the concentration and
    delivering method with inhibitor

Assimilation of lutein
  1. Decrease of 70 (mean value)
  2. Saturation versus linear trend
  3. Increase of efficiency at 0.5 mM

18
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Primary effects of the factors concentration,
    donor solution type and inhibition

19
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • SUMMARY
  • Factors concentration, donor solution type and
    inhibition
  • Assimilation of carotenoids from micellar
    solutions is significantly inhibited with the use
    of HDLs
  • Significant decrease of the assimilation level,
    (70 drop for lutein), although it did not
    reached 100. Co-existence of simple diffusion
    mechanism and work of transporters not totally
    blocked under the established experimental
    conditions
  • Carotenes were more efficiently absorbed than
    lutein even under inhibition conditions. They
    probably take help of different protein
    transporters

20
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • SUMMARY
  • Factors concentration, donor solution type and
    inhibition
  • Carotenoid-CyDIC were more efficiently absorbed
    than the carotenoid micellar solutions under
    inhibition conditions. How did the factor
    inhibition affect the carotenoid assimilation
    from CyDIC?

21
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotene-CyDIC absorption
    efficiency in function of the inhibition factor

Assimilation of b-carotene-CyDIC
  • Increase of efficiency from 0.5 mM
  • under inhibited transport conditions
  • Increase of 86 at 1.0 mM

22
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotene-CyDIC absorption
    efficiency in function of the inhibition factor

Assimilation of lycopene-CyDIC
  • Increase of efficiency from 0.5 mM
  • under inhibited transport conditions
  • Increase of 165 at 1.0 mM

23
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotene-CyDIC absorption
    efficiency in function of the inhibition factor

24
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the carotene-CyDIC absorption
    efficiency in function of the inhibition factor

25
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • RESULTS
  • Comparison of the lutein absorption efficiency in
    function of the inhibition factor

Assimilation of lutein at 1.0 mM
  • 70 drop of micellar lutein
  • under inhibited transport conditions
  • 28 drop of lutein-CyDIC under
  • inhibited transport conditions

26
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • SUMMARY
  • Comparison of the carotenoid-CyDIC absorption
    efficiency in function of the inhibition factor
  • A different effect of HDLs was observed for the
    assimilation efficiency of carotene-CyDICs or
    lutein-CyDICs
  • Process of competition between HDLs and
    lutein-CyDIC may not be efficient enough in
    comparison with the same process for
    carotene-CyDIC
  • Inhibition promoted by HDLs affects specific
    protein transporters

27
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • SUMMARY
  • Comparison of the in vivo lutein absorption
    efficiency non-responder versus
    lutein-responders group

non-Responders group
Lutein-responders group
28
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • CONCLUSIONS
  • Factors concentration, donor solution type and
    inhibition
  • First, inter-individual differences on carotenoid
    assimilation efficiency should be evaluated, as
    they are a direct consequence of facilitated
    diffusion mechanism and expression/location of
    transporters. Interaction with drugs
  • New strategies to increase carotenoid
    assimilation to develop food formulae.
    Interaction with lipoprotein/apoprotein components

29
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • CONCLUSIONS
  • Factors concentration, donor solution type and
    inhibition
  • Data point to the existence of different affinity
    of transporters and even different transporters
    for carotenes and the xanthophyll lutein.
    Non-/low-responder effect
  • Bringing pharmaceutical concepts to food
    technology and nutrition will help to consolidate
    functional food

30
In vitro intestinal absorption of carotenoids
delivered as molecular inclusion complexes
  • ACKNOWLEGMENTS
  • Financial support from Spanish Government
    (projects AGL2007-61146 AGR-03025)
  • Scientific and organizing committees of the 6th
    International Congress on Pigments in Food -
    Budapest

Dr. Antonio Pérez-Gálvez aperez_at_cica.es
Food Biotechnology Department Instituto de la
Grasa (CSIC) Av. Padre García Tejero 4, 41012
Sevilla (SPAIN)
THANKS FOR YOUR ATTENTION!
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