Association of bioplymers in microencapsulation to control the release of active

molecules

Laura Snchez-Gonzlez*, Ghazi Ben Messaoud, Stphane Desobry

Laboratoire dIngnierie des Biomolcules (LIBio). ENSAIA - Universit de Lorraine.

2 avenue de la Fort de Haye, TSA 40602, 54518 Vanduvre-ls-Nancy Cedex, France.

Abstract

Alginate, the major structural component of the brown seaweeds and bacterial capsules,

is one of the most used biopolymers in encapsulation. Among the developed systems,

liquid-core capsules with alginate membrane have a wide range of applications in

different areas such as food, nutraceuticals and biomedical. To improve the

physicochemical properties of these systems and in particular the barrier properties an

interesting approach involves the incorporation of a second polymer. Two examples

were presented: (a) composite capsules with a membrane of sodium caseinate / alginate

which improved stability and pH-dependent release of a dye molecule used as a model

(b) incorporation of shellac in the membrane or as an external coating to enhance the

barrier properties for low molecular weight molecules (e.g. riboflavin).

Mots-cls : biopolymers, alginate hydrogel, liquid-core capsules, permeability, release

pH-dependent.

(*) Contact information for Corresponding Author

Phone: +33 (0)3 83 59 58 77; Fax: +33 (0)3 83 59 58 04; e-mail: laura.sanchez-

gonzalez@univ-lorraine.fr

1
1. Encapsulation in an alginate matrix

Alginate is a biopolymer widely used in industry because it has many properties:

thickener, stabilizer, gelling and film-forming property and is abundant in nature.

Indeed, it is a major structural component of brown algae (up to 40% of the total dry

weight) and bacterial capsules. Sale of alginates began in 1927, and world production

reached today 30,000 tons / year. About 30 % of this production is for the food industry

and the rest of the production is distributed over several sectors, particularly the

pharmaceutical and cosmetic industry [1].

1.1. Chemical structure and gelation

Alginate is a copolymer which consists of alternating -D mannuronic acid (M) and -L

guluronic acid (G) linked in (14). Uronic acid molecules are distributed along the

chain of alginate in the form of blocks. There are 3 types: homopolymeric sequences

containing only mannuronic acid units (M units) or guluronic (G blocks), and

heteropolymeric blocks containing both mannuronic and guluronic acid units (MG

blocks) [2]. The composition and organization of the uronic acid sequences are highly

variable depending on the type of alginate. Concerning the chains conformation, four

glycosidic bonds were present in alginate: di-axial (GG), di-equatorial (MM), axial-

equatorial (GM) axial and equatorial (MG).

The ionotropic gelation of the alginate is related to its affinity for certain ions and the

formation of selective bonds [3,4]. Indeed, alginate polymer chains interact with

divalent or trivalent cations (Fe3+, Al3+) to form hydrogels. The affinity of the alginate

against divalent ions decreases in the following order: Pb> Cu> Cd> Ba> Sr> Ca> Co,

Ni, Zn> Mn [5]. Calcium remains the most commonly cation used to induce alginate gel

2
formation. Two G or MG blocks of two adjacent polymer chains may be crosslinked in

the presence of a divalent cation by electrostatic interaction between carboxyl groups of

the polysaccharide COO-. When two polymer strands are facing each other, the

conformation of G or MG blocks form cavities in which are housed the Ca2 + ions. This

blocks dimerization is named "egg box" structure. [6]

The ionotropic gelation is the most used method for the formation of alginate gels.

Indeed, it is simple to implement gelation under relatively mild conditions. These gels

are stable in neutral or slightly acidic conditions and are modified into acid gels by

proton exchange at pH <3-3.5 (pka alginate). However, under basic solutions, alginate

gels undergo swelling followed by an erosion and possibly disintegration. Control of

physicochemical properties of this type of biomaterial (mechanical properties, swelling,

degradation ..) is also limited by the loss of divalent cations in the surrounding medium

by ion exchange which affects the chemical stability of the matrix. For instance, an

alginate hydrogel degrades in the presence of saline solutions [7]. In this context,

several research teams have focused on the preparation of gels using other crosslinking

methods after chemical modification of alginate as covalent crosslinking or

photocrosslinking.

1.2. Alginate hydrogels applications

Alginate hydrogel represent a type of biomaterials with a wide range of applications in

many fields such as food [8], nutraceutical [9], environment and water treatment [10].

The interest of alginate hydrogels also gave rise to the development of a wide range of

biomaterials with different architectures such as plane or spherical hydrogels (beads and

capsules), fibers and anisotropic structures at milli -, micro- and nanoscale (Figure 1).

3
1.3. Encapsulations techniques using alginate

Several encapsulation techniques using alginate were developed to obtain particles with

a wide size range. Indeed, the diameter of the alginate gel particles can be> 1000

microns (macro), from 0.2 to 1000 microns (micro) or <0.2 microns (nanoparticles).

Table 1 summarizes the encapsulation techniques using alginate, the particle size

obtained and the potential applications of different systems [17].

1.4. Alginate liquid-core microcapsules

The liquid-core capsules with a calcium alginate hydrogel membrane can be prepared

according to three main approaches that are the electro-coextrusion and coextrusion, the

multilayer polymerization and the inverse spherification.

The first approach is the use of minifluidic or millifluidic instruments, by coextrusion or

by electro co-extrusion of the core of the capsule (internal mixing) and the alginate

solution (outer solution) in a bath of CaCl2 to form the alginate capsules. The thickness

of the membrane is controlled by changing the ratio between the flow rates of the inner

and outer solutions [18].

The multilayer polymerization is based on the formation of alginate beads by simple

extrusion of an alginate solution into a bath of CaCl2. Spherical alginate hydrogels are

then coated by the addition of a layer of cationic polymer or more coating layers

alternating alginate with an opposite charged polymer as chitosan or Poly-L-lysine. The

coating operation may be repeated until the desired number of layers [19]. Finally, the

core alginate was dissolved by incubating the capsules in a solution of chelating agent

(sodium citrate, EDTA).

Concerning the reverse gelation, the CaCl2 droplets containing the active molecule are

injected into a bath of sodium alginate, an alginate membrane formed instantly when the

4
two solutions come into contact, partially gelled capsules are then transferred into a

CaCl2 solution to complete the crosslinking process. The thickness of the membrane

depends on the sodium alginate concentration and contact time in the polymer solution.

After the gelling bath, the capsules should be washed in order to prevent aggregation of

capsules by external gelation [20].

2. Improvement of physico-chemical properties of alginate capsules by polymers

association

The major limitation to the use of alginate liquid-core capsules is the significant and

rapid loss of their contents. The physico-chemical properties (membrane permeability,

mechanical properties ..) of capsules may be improved by the incorporation of a second

polymer.

In this approach two examples will be presented: the incorporation of sodium caseinate

in the alginate membrane and shellac incorporation in the membrane of the capsules or

as an external coating. The association of two biopolymers enables the development of

pH-dependent release systems.

2.1. Incorporation of a polymer in the membrane which can interact with the

encapsulated molecule, sodium caseinate [21].

In order to control the diffusion of the molecule encapsulated in alginate capsules, an

interesting strategy is the incorporation of a second polymer in the membrane. The

selected polymer (sodium caseinate) has the ability to interact with the encapsulated

molecule under specific environmental conditions (pH) and thus limit its release.

The composite capsules showed a strengthening of the mechanical properties which can

be explained by the interactions between the carboxylic groups of the protein mixture

5
and the calcium cations, but also by the electrostatic interactions between the carboxyl

groups of the alginate and the amine groups of the caseinate sodium. The diffusion

kinetics of cochineal red (used as a model molecule in this study) showed a pH-

dependent release profile (Figure 2). Composite capsules displayed a retarded release

with high dye retention at pH 2 and 3 compared to NaAlg capsules. By adding NaCas

into alginate membrane, the amount of dye released after 2h was reduced from 95 %

to 7 % at pH 2. This controlled release is explained by a membrane dye adsorption

thanks to the establishment of electrostatic interactions between the sulphonate and

amines groups of caseinate.

2.2. Incorporation of a pH-sensitive polymer, shellac [22].

In order to improve the barrier properties of the liquid-core capsules, an interesting

approach is the combination of two polymers, alginate of the membrane and a pH-

sensitive polymer: shellac (E904) from the secretion of an Asian scale insect (Kerria

lacca). For this, the shellac has been incorporated directly to the membrane of the

capsules (composite system) or as outer layer (coated system) and these two systems

were compared with respect to a control (alginate membrane). The results showed

significant change in physico-chemical properties of the alginate capsules with shellac.

The release kinetics of riboflavin used as model molecule was studied for three systems

(Figure 3). A pH-dependent release of the low molecular weight molecule was observed

for the coated system in the pH range tested. Indeed the total amount of riboflavin

released decreased from 90 % (control and composite system) to 52 and 60 % with

coated system at pH 1 and 3 respectively.

References

6
[1] Ertesvg, H., & Valla, S. (1998). Biosynthesis and applications of alginates.
Polymer Degradation and Stability, 59(13), 8591.
[2] Pawar, S. N., & Edgar, K. J. (2012). Alginate derivatization: A review of chemistry,
properties and applications. Biomaterials, 33(11), 32793305.
[3] Haug, A., Smidsrd, O., Hgdahl, B., ye, H. A., Rasmussen, S. E., Sunde, E., &
Srensen, N. A. (1970). Selectivity of Some Anionic Polymers for Divalent Metal Ions.
Acta Chemica Scandinavica, 24, 843854.
[4] Smidsrd, O. (1974). Molecular basis for some physical properties of alginates in
the gel state. Faraday Discussions of the Chemical Society, 57(0), 263274.
[5] Mrch, Y. A., Donati, I., Strand, B. L., & Skjk-Braek, G. (2006). Effect of Ca2+,
Ba2+, and Sr2+ on alginate microbeads. Biomacromolecules, 7(5), 14711480.
[6] Grant, G. T., Morris, E. R., Rees, D. A., Smith, P. J., & Thom, D. (1973). Biological
interactions between polysaccharides and divalent cations: the egg-box model. FEBS
Letters, 32(1), 195198.
[7] Rolland, L., Santanach-Carreras, E., Delmas, T., Bibette, J., & Bremond, N. (2014).
Physicochemical properties of aqueous core hydrogel capsules. Soft Matter, 10(48),
96689674.
[8] Farris, S., Schaich, K. M., Liu, L., Piergiovanni, L., & Yam, K. L. (2009).
Development of polyion-complex hydrogels as an alternative approach for the
production of bio-based polymers for food packaging applications: a review. Trends in
Food Science & Technology, 20(8), 316332.
[9] Chen, L., Remondetto, G. E., & Subirade, M. (2006). Food protein-based materials
as nutraceutical delivery systems. Trends in Food Science & Technology, 17(5), 272
283.
[10] Crini, G. (2005). Recent developments in polysaccharide-based materials used as
adsorbents in wastewater treatment. Progress in Polymer Science, 30(1), 3870.
[11] Bremond, N., Santanach-Carreras, E., Chu, L.-Y., & Bibette, J. (2010). Formation
of liquidcore capsules having a thin hydrogel membrane: liquid pearls. Soft Matter,
6(11), 24842488.
[12] Zhu, H., Srivastava, R., Brown, J. Q., & McShane, M. J. (2005). Combined
physical and chemical immobilization of glucose oxidase in alginate microspheres
improves stability of encapsulation and activity. Bioconjugate Chemistry, 16(6), 1451
1458.
[13] Lertsutthiwong, P., Noomun, K., Jongaroonngamsang, N., Rojsitthisak, P., &
Nimmannit, U. (2008). Preparation of alginate nanocapsules containing turmeric oil.
Carbohydrate Polymers, 74(2), 209214.
[14] Mele, E., Fragouli, D., Ruffilli, R., Gregorio, G. L. D., Cingolani, R., &
Athanassiou, A. (2013). Complex architectures formed by alginate drops floating on
liquid surfaces. Soft Matter, 9(27), 63386343.
[15] Shin, S.-J., Park, J.-Y., Lee, J.-Y., Park, H., Park, Y.-D., Lee, K.-B., Lee, S.-H.
(2007). On the Fly Continuous Generation of Alginate Fibers Using a Microfluidic
Device. Langmuir, 23(17), 91049108.
[16] Lee, K. H., Shin, S. J., Park, Y., & Lee, S.-H. (2009). Synthesis of Cell-Laden
Alginate Hollow Fibers Using Microfluidic Chips and Microvascularized Tissue-
Engineering Applications. Small, 5(11), 12641268.
[17] Ching, S. H., Bansal, N., & Bhandari, B. (2015). Alginate gel particles- a review of
production techniques and physical properties. Critical Reviews in Food Science and
Nutrition, 0.

7
[18] Bremond, N., Santanach-Carreras, E., Chu, L.-Y., & Bibette, J. (2010). Formation
of liquidcore capsules having a thin hydrogel membrane: liquid pearls. Soft Matter,
6(11), 24842488.
[19] Breguet, V., Gugerli, R., Pernetti, M., von Stockar, U., & Marison, I. W. (2005).
Formation of Microcapsules from Polyelectrolyte and Covalent Interactions. Langmuir,
21(21), 97649772.
[20] Blandino, A., Macas, M., & Cantero, D. (1999). Formation of calcium alginate gel
capsules: influence of sodium alginate and CaCl2 concentration on gelation kinetics.
Journal of Bioscience and Bioengineering, 88(6), 686689.
[21] Ben Messaoud, G., Snchez-Gonzlez, L., Jacquot, A., Probst, L., & Desobry, S.
(2015). Alginate/sodium caseinate aqueous-core capsules: a pH-responsive matrix.
Journal of Colloid and Interface Science, 440, 1-8.
[22] Ben Messaoud, G., Snchez-Gonzlez, L., Probst, L., Degen, P., & Desobry. S.
Physicochemical properties of alginate/shellac aqueous-core capsules: influence of
membrane architecture. (submitted)

8
Tableau 1. Synthesis of the current methods of production of particles based on
alginate gel.

Techniques Size particles Applications

Simple extrusion 1000-2000 m Encapsulation of probiotics, yeasts, fish

oil, medicines, proteins, enzymes, plant
extracts and flavors

Electrostatic potential 0.9-1.5 m Encapsulation of lipid nanoparticles,

50-350 m probiotics, animal and human tissues and
enzymes

Vibrating nozzle <20 m Encapsulation of cells and proteins

200-5000 m

Jet broken 200- 5000 m Encapsulation of cells and proteins

Rotating disc 300-600 m Encapsulation of bacteria, yeasts and

proteins

Rotating nozzle 270-2000 m Encapsulation of cells and proteins

Spray nozzle 80-130 m Encapsulation of lipidic nanoparticles,

lipidiques, BCG vaccine, antigens,
proteins and bacteria

Spray (against the 11-47 m Encapsulation of fish oil, lysozyme,

current) ibuprofen, propranolol hydrochloride,
Impinging aerosol probiotics, insulin

Emulsification 100- 1000 m Encapsulation of insulin

Microfluidic 50 - 70 m Encapsulation of cells (probiotics,
mammals)

Micro- nano-molding 120 200 nm Encapsulation of proteins (BSA)

Hybrid microgels 250-600 nm Encapsulation of drugs (doxorubicin),

cells (hemoglobin), insulin and herbicides

9
 Figure 1. Example of biomaterials developed from alginate hydrogel. Millimetric
liquid-core capsules with a thin membrane of alginate (a) alginate microspheres
obtained by emulsification (b) alginate nanocapsules (c) image of anisotropic capsules
mushroom-shaped (d), full microfiber (e) hollow microfibers (f) [11,12,13,14,15,16]

10
 ! NaAlg capsules NaAlg-NaCas (1:0.5)

100 100

80 80

60 60

40 40

Release (%)
Release (%)

20 20

0 0
0 100 200 300 400 0 100 200 300 400
Time (min) Time (min)

NaAlg-NaCas (1:1) NaAlg-NaCas (1:1.5)

100 100

80 80

60 60

40 40
Release (%)
Release (%)

20 20

0 0
0 100 200 300 400 0 100 200 300 400
Time (min) Time (min)

Figure 2. Cochineal red dye release for alginate and composite capsules at pH 2 (), 3
( ), 4.8 ( ) and 7 ( ). The solid lines represent the results of fits of the experimental
data.[21].

11
 pH1

100

80

60

40

Release (%)
Simple
20 Composite
coat 1m in vs Col 8
coat 10m in vs Col 11

0
0 100 200 300
Time (min)

pH 3

100

80

60
Release (%)

40

Simple
Composite
20
Coated 1 min
Coated 10 min
0
0 100 200 300
Time (min)

pH 5

100

80

60

40
Release (%)

Simple
Composite
20 Coated 1 min
Coated 10 min

0
0 100 200 300
Time (min)

Figure 3. Riboflavin release from simple alginate, alginate-shellac gum composite and
coated capsules during 1 and 10 min via shellac gum acid precipitation at different pH-
values [22].

12
Association de biopolymres en microencapsulation en vue de matriser la

libration de composs d'intrt

Association of bioplymers in microencapsulation to control release of active

molecules

Laura Snchez-Gonzlez*, Ghazi Ben Messaoud, Stphane Desobry

Laboratoire dIngnierie des Biomolcules (LIBio). ENSAIA - Universit de Lorraine.

2 avenue de la Fort de Haye, TSA 40602, 54518 Vanduvre-ls-Nancy Cedex, France.

Rsum

Lalginate, compos structural majeur des algues brunes et des capsules bactriennes est

un des biopolymres les plus utiliss en encapsulation. Parmi les systmes dvelopps,

les capsules cur liquide avec une membrane dalginate ont un large spectre

dapplications dans des domaines tels que lalimentaire, la nutraceutique ou le

biomdical. Pour amliorer les proprits physico-chimiques de ces systmes et

notamment les proprits barrire une approche intressante consiste en lincorporation

dun deuxime polymre. Deux exemples sont prsents : (a) des capsules composites

avec une membrane de casinate de sodium/alginate qui prsentent une meilleure stabilit et une

libration pH-dpendante dun colorant utilis comme molcule modle (b) lincorporation de la

gomme laque dans la membrane ou comme revtement externe pour amliorer les proprits

barrire vis--vis de molcules de faible masse molculaire (riboflavine par exemple).

Mots-cls : biopolymres, hydrogel dalginate, capsules cur liquide, permabilit,

libration pH-dpendant.

(*) Contact information for Corresponding Author

Phone: +33 (0)3 83 59 58 77; Fax: +33 (0)3 83 59 58 04; e-mail: laura.sanchez-

gonzalez@univ-lorraine.fr

13
1. Encapsulation dans une matrice dalginate

Lalginate est un biopolymre polyvalent trs utilis en industrie car il possde de

nombreuses proprits: paississant, stabilisant, glifiant et pouvoir filmogne et il est

abondant dans la nature. En effet, il sagit dun compos structural majeur des algues

brunes (jusqu 40% de la masse sche totale) et des capsules bactriennes. La

commercialisation des alginates a dbut en 1927, et la production mondiale atteint de

nos jours 30 000 tonnes/an. Environ 30 % de cette production est destine lindustrie

agroalimentaire et le reste de la production est repartie sur plusieurs secteurs et plus

particulirement lindustrie pharmaceutique et cosmtique [1].

1.1. Structure chimique et glification

Lalginate est un copolymre qui consiste en une alternance dacide -D mannuronique

(M) et dacide -L guluronique (G) lis en (14). Les molcules dacide uronique sont

rparties le long de la chane de lalginate sous forme de blocs. Il en existe 3 types: des

squences homopolymriques ne contenant que des units dacide mannuronique (blocs

M) ou guluronique (blocs G), ainsi que des blocs htropolymriques contenant la fois

des units dacide mannuronique et guluronique (blocs MG) [2]. La composition et

lorganisation des acides uroniques en squences sont donc trs variables en fonction du

type dalginate. En ce qui concerne la conformation des chaines, lalginate possde

quatre types de liaisons glycosidiques : di-axial (GG), di-quatoriale (MM), axial-

quatoriale (GM) et quatoriale axiale (MG).

La glification ionotropique de lalginate est lie son affinit pour certains ions et la

formation de liaisons slectives [3,4]. En effet, les chaines de polymre dalginate

interagissent avec les cations bivalents ou trivalents (Fe3+, Al3+) pour former des

14
hydrogels. L'affinit de lalginate vis--vis des ions bivalents diminue dans l'ordre

suivant: Pb> Cu> Cd> Ba> Sr> Ca> Co, Ni, Zn> Mn [5]. Le calcium demeure le cation

le plus couramment utilis pour induire la formation de gel d'alginate. Deux blocs G ou

MG de deux chanes de polymres adjacentes peuvent tre rticuls en prsence dun

cation bivalent par interaction lectrostatique entre deux groupements carboxyliques

COO du polysaccharide. Lorsque deux brins de polymre sont face face, la

conformation des blocs G ou MG forment des cavits dans lesquelles viennent se loger

les ions Ca2+. Cette dimrisation des blocs est appele structure type "boite dufs" [6].

La glification ionotropique est la mthode la plus utilise pour la formation des gels

dalginate. En effet, cest une glification simple mettre en oeuvre dans des conditions

relativement douces. Ces gels sont stables dans des conditions de pH neutre ou

lgrement acide et sont convertis en gels acides par change de protons des pH acides

< 3-3.5 (pka de lalginate). Cependant, dans des solutions faiblement basiques, les gels

dalginate subissent un gonflement suivis par une rosion et ventuellement une

dsintgration. Le contrle des proprits physicochimiques de ce type de biomatriau

(proprits mcanique, gonflement, dgradation..) est galement limit par la perte de

cations bivalents dans le milieu environnant par change dions ce qui affecte la stabilit

chimique de la matrice. Par exemple, un hydrogel d'alginate se dgrade en prsence de

solutions salines [7]. Dans ce contexte, plusieurs quipes se sont intresses la

prparation de gels en utilisant dautres mthodes de rticulation aprs modification

chimique de lalginate comme la rticulation covalente ou la photorticulation de

lalginate.

1.2. Applications des hydrogels dalginate

15
Les hydrogels dalginate constituent une classe de biomatriaux avec un large spectre

dapplications dans plusieurs domaines, lalimentaire [8], nutraceutique [9],

environnement et traitement des eaux [10]. Lintrt des hydrogels base dalginate a

galement donn naissance au dveloppement dune large gamme de biomatriaux avec

diffrentes architectures tels que des hydrogels plan ou sphriques (billes et capsules),

de fibres et des structures anisotropes lchelle milli-, micro- et nanomtriques (Figure

1).

1.3. Techniques encapsulation avec de lalginate

Plusieurs techniques dencapsulation en utilisant lalginate ont t dveloppes

permettant dobtenir des particules avec une large gamme de taille. En effet, le diamtre

de particules de gel d'alginate peut tre > 1000 m (macro), de 0,2 1000 m (micro)

ou <0,2 m (nanoparticules). Le tableau 1 rsume les techniques dencapsulation

utilisant lalginate, la taille des particules obtenue ainsi que les applications potentielles

des diffrents systmes [17].

1.4. Microcapsules dalginate cur liquide

Les capsules coeur liquide prsentant une membrane dhydrogel dalginate de calcium

peuvent tre prpares selon trois principales approches qui sont la coextrusion et

electro-coextrusion, la polymrisation multicouches et la sphrification inverse.

La premire approche consiste en lutilisation de dispositifs de millifluidique ou

minifluidique, par co-extrusion ou par lectro co-extrusion du cur de la capsule

(mlange interne) et de la solution dalginate (solution priphrique) dans un bain de

CaCl2 pour former les capsules d'alginate Lpaisseur de la membrane est contrle en

changeant le rapport entre les dbits de la solution interne et externe [18].

16
La polymrisation multicouches est base sur la formation de billes d'alginate par

simple extrusion goutte goutte dune solution dalginate dans un bain de CaCl2. Les

hydrogels sphriques dalginate sont ensuite enrobs par lajout dune couche de

polymre cationique ou de plusieurs couches de revtement en alternant lalginate avec

un polymre de charges oppose de type chitosane ou Poly-L-lysine. Lopration

denrobage peut tre rpte jusqu lobtention de nombre de couches souhait [19].

Enfin, le cur dalginate est dissout par incubation des capsules dans une solution

dagent chlatant (citrate de sodium, EDTA).

Concernant la glification inverse, les gouttelettes de CaCl2 contenant la molcule

active sont injectes goutte goutte dans un bain d'alginate de sodium, une membrane

dalginate se forme instantanment lorsque les deux solutions entrent en contact, aprs

un certain temps de sjour, les capsules partiellement glifis sont transfres dans une

solution de CaCl2 pour terminer le processus de rticulation. Lpaisseur de la

membrane dpend de la concentration en alginate de sodium et du temps de sjour dans

la solution de polymre. A la sortie du bain de glification, les capsules doivent tre

rinces afin dviter lagrgation des capsules par glification externe [20].

2. Amlioration des proprits physico-chimiques des capsules dalginate cur

liquide par incorporation de biopolymre

Une des principales limites lutilisation des capsules dalginate cur liquide est la

perte significative et rapide de leur contenu. Les proprits physico-chimiques

(permabilit de la membrane, proprits mcaniques..) des capsules peuvent tre

amliores par incorporation dun deuxime polymre.

Suivant cette approche deux exemples seront prsents : lincorporation de caseinate de

sodium dans la membrane dalginate et lincorporation de gomme laque dans la

17
membrane des capsules ou comme enrobage externe. Lassociation de deux

biopolymres permet de dvelopper des systmes de libration pH-dpendant.

2.1. Incorporation dun polymre dans la membrane pouvant interagir avec la

molcule encapsule, le casinate de sodium [21].

Dans le but de contrler la diffusion de la molcule encapsule dans les capsules base

dalginate, une stratgie intressante consiste en lincorporation dun second polymre

dans la membrane. Le polymre slectionn (caseinate de sodium) a la capacit

dinteragir avec la molcule encapsule dans des conditions environnementales

spcifiques (pH) et donc de limiter ainsi sa libration.

Les capsules composites ont montr un renforcement des proprits mcaniques qui

peut tre expliqu par des interactions entre les groupements carboxyliques du mlange

protique et les cations calcium, mais aussi par les interactions lectrostatiques entre les

groupements carboxyliques de lalginate et les groupements amines du caseinate de

sodium. La cintique de diffusion du rouge cochenille (utilis comme molcule modle

dans cette tude) a montr un profil de libration pH-dpendant (Figure 2). La libration

du colorant partir des capsules composites se fait avec un retard par rapport aux

capsules dalginate et une forte rtention du colorant est observe pH 2 et 3. Suite

lincorporation de caseinate de sodium dans la membrane dalginate, la quantit de

colorant libre aprs 2 heures a t rduite de 95 7 % pH 2. Cette libration

contrle est explique par une adsorption membranaire du colorant par des interactions

lectrostatiques entre les groupements sulfonates et amines du caseinate.

2.2. Incorporation dun polymre pH-sensible, la gomme laque [22].

18
Dans le but damliorer les proprits barrire des capsules cur liquide, une

approche intressante consiste en lassociation de deux polymres, lalginate de la

membrane et un polymre pH-sensible: la gomme laque (E904) issue de scrtion dune

cochenille asiatique (Kerria lacca). Pour cela, la gomme laque a t incorpore soit

directement au niveau de la membrane des capsules (systme composite) soit applique

comme couche externe (systme enrob) et ces deux systmes ont t compars par

rapport un systme alginate (tmoin). Les rsultats ont montr une modification

significative des proprits physico-chimiques des capsules suite lutilisation de

gomme laque. La cintique de libration de la riboflavine utilise comme molcule

modle a t tudie pour les 3 systmes (Figure 3). Une libration pH-dpendante de la

molcule tmoin de faible poids molculaire a t observe pour le systme enrob dans

lintervalle de pH test. En effet, la quantit maximale de riboflavine libre passe de

90 % (tmoin et composite) 52 et 60 % pour le systme enrob pH 1 et 3

respectivement.

Rfrences bibliographiques

[1] Ertesvg, H., & Valla, S. (1998). Biosynthesis and applications of alginates.
Polymer Degradation and Stability, 59(13), 8591.
[2] Pawar, S. N., & Edgar, K. J. (2012). Alginate derivatization: A review of chemistry,
properties and applications. Biomaterials, 33(11), 32793305.
[3] Haug, A., Smidsrd, O., Hgdahl, B., ye, H. A., Rasmussen, S. E., Sunde, E., &
Srensen, N. A. (1970). Selectivity of Some Anionic Polymers for Divalent Metal Ions.
Acta Chemica Scandinavica, 24, 843854.
[4] Smidsrd, O. (1974). Molecular basis for some physical properties of alginates in
the gel state. Faraday Discussions of the Chemical Society, 57(0), 263274.
[5] Mrch, Y. A., Donati, I., Strand, B. L., & Skjk-Braek, G. (2006). Effect of Ca2+,
Ba2+, and Sr2+ on alginate microbeads. Biomacromolecules, 7(5), 14711480.
[6] Grant, G. T., Morris, E. R., Rees, D. A., Smith, P. J., & Thom, D. (1973). Biological
interactions between polysaccharides and divalent cations: the egg-box model. FEBS
Letters, 32(1), 195198.
[7] Rolland, L., Santanach-Carreras, E., Delmas, T., Bibette, J., & Bremond, N. (2014).
Physicochemical properties of aqueous core hydrogel capsules. Soft Matter, 10(48),
96689674.

19
[8] Farris, S., Schaich, K. M., Liu, L., Piergiovanni, L., & Yam, K. L. (2009).
Development of polyion-complex hydrogels as an alternative approach for the
production of bio-based polymers for food packaging applications: a review. Trends in
Food Science & Technology, 20(8), 316332.
[9] Chen, L., Remondetto, G. E., & Subirade, M. (2006). Food protein-based materials
as nutraceutical delivery systems. Trends in Food Science & Technology, 17(5), 272
283.
[10] Crini, G. (2005). Recent developments in polysaccharide-based materials used as
adsorbents in wastewater treatment. Progress in Polymer Science, 30(1), 3870.
[11] Bremond, N., Santanach-Carreras, E., Chu, L.-Y., & Bibette, J. (2010). Formation
of liquidcore capsules having a thin hydrogel membrane: liquid pearls. Soft Matter,
6(11), 24842488.
[12] Zhu, H., Srivastava, R., Brown, J. Q., & McShane, M. J. (2005). Combined
physical and chemical immobilization of glucose oxidase in alginate microspheres
improves stability of encapsulation and activity. Bioconjugate Chemistry, 16(6), 1451
1458.
[13] Lertsutthiwong, P., Noomun, K., Jongaroonngamsang, N., Rojsitthisak, P., &
Nimmannit, U. (2008). Preparation of alginate nanocapsules containing turmeric oil.
Carbohydrate Polymers, 74(2), 209214.
[14] Mele, E., Fragouli, D., Ruffilli, R., Gregorio, G. L. D., Cingolani, R., &
Athanassiou, A. (2013). Complex architectures formed by alginate drops floating on
liquid surfaces. Soft Matter, 9(27), 63386343.
[15] Shin, S.-J., Park, J.-Y., Lee, J.-Y., Park, H., Park, Y.-D., Lee, K.-B., Lee, S.-H.
(2007). On the Fly Continuous Generation of Alginate Fibers Using a Microfluidic
Device. Langmuir, 23(17), 91049108.
[16] Lee, K. H., Shin, S. J., Park, Y., & Lee, S.-H. (2009). Synthesis of Cell-Laden
Alginate Hollow Fibers Using Microfluidic Chips and Microvascularized Tissue-
Engineering Applications. Small, 5(11), 12641268.
[17] Ching, S. H., Bansal, N., & Bhandari, B. (2015). Alginate gel particles- a review of
production techniques and physical properties. Critical Reviews in Food Science and
Nutrition, 0.
[18] Bremond, N., Santanach-Carreras, E., Chu, L.-Y., & Bibette, J. (2010). Formation
of liquidcore capsules having a thin hydrogel membrane: liquid pearls. Soft Matter,
6(11), 24842488.
[19] Breguet, V., Gugerli, R., Pernetti, M., von Stockar, U., & Marison, I. W. (2005).
Formation of Microcapsules from Polyelectrolyte and Covalent Interactions. Langmuir,
21(21), 97649772.
[20] Blandino, A., Macas, M., & Cantero, D. (1999). Formation of calcium alginate gel
capsules: influence of sodium alginate and CaCl2 concentration on gelation kinetics.
Journal of Bioscience and Bioengineering, 88(6), 686689.
[21] Ben Messaoud, G., Snchez-Gonzlez, L., Jacquot, A., Probst, L., & Desobry, S.
(2015). Alginate/sodium caseinate aqueous-core capsules: a pH-responsive matrix.
Journal of Colloid and Interface Science, 440, 1-8.
[22] Ben Messaoud, G., Snchez-Gonzlez, L., Probst, L., Degen, P., & Desobry. S.
Physicochemical properties of alginate/shellac aqueous-core capsules: influence of
membrane architecture. (submitted)

20
Tableau 1. Synthse des mthodes actuelles de production de particules base de gel
d'alginate.

Techniques Taille des particules Applications

Encapsulation des probiotiques, des
Simple extrusion levures, huile de poisson, des
1000-2000 m mdicaments, des protines, des enzymes,
des extraits de plantes et saveurs

Potentiel 0.9-1.5 m Encapsulation de nanoparticules lipidiques,

lectrostatique 50-350 m des probiotiques, des tissus animaux et
humains et des enzymes.

Buse vibrante <20 m Encapsulation de cellules et de protines

200-5000 m

Jet rompu 200- 5000 m Encapsulation de cellules et de protines

Disque tournant 300-600 m Encapsulation des bactries, de la levure et

les protines

Buse tournante 270-2000 m Encapsulation de cellules et de protines

Buse de pulvrisation 80-130 m Encapsulation de nanoparticules lipidiques,

vaccin BCG, des antignes, des protines
et des bactries.

Atomisation ( contre- 11-47 m Encapsulation de l'huile de poisson, le

courant) lysozyme, l'ibuprofne, le chlorhydrate de
Impinging aerosol propranolol, les probiotiques, l'insuline

Emulsification 100- 1000 m Encapsulation de linsuline

Microfluidique 50 - 70 m Encapsulation de cellules (probiotiques,
mammifres)

Micro- nano-moulage 120 200 nm Encapsulation de protines (BSA)

Microgels hybrides 250-600 nm Encapsulation de mdicaments

(doxorubicine), cellules (hmoglobine),
l'insuline et herbicides

21
 Figure 1. Exemple de biomatriaux dvelopps partir dun hydrogel dalginate.
Capsules millimtrique coeur liquide avec une fine membrane
dalginate(a),microsphres dalginate obtenues par mulsification (b), nanocapsules
dalginate (c), photo de capsules anisotrope en forme de champignon (d), microfibre
pleine (e) et microfibres creuse (f). [11,12,13,14,15,16]

22
 ! NaAlg capsules NaAlg-NaCas (1:0.5)

100 100

80 80

60 60

40 40

Release (%)
Release (%)

20 20

0 0
0 100 200 300 400 0 100 200 300 400
Time (min) Time (min)

NaAlg-NaCas (1:1) NaAlg-NaCas (1:1.5)

100 100

80 80

60 60

40 40
Release (%)
Release (%)

20 20

0 0
0 100 200 300 400 0 100 200 300 400
Time (min) Time (min)

Figure 2. Cintique de libration du colorant rouge cochenille encapsul dans des

capsules dalginate ou composites diffrents pH 2 (), 3 ( ), 4.8 ( ) and 7 ( ). Les
lignes continues sont le rsultat de la modlisation des donnes exprimentales. [21].

23
 pH1

100

80

60

40

Release (%)
Simple
20 Composite
coat 1m in vs Col 8
coat 10m in vs Col 11

0
0 100 200 300
Time (min)

pH 3

100

80

60
Release (%)

40

Simple
Composite
20
Coated 1 min
Coated 10 min
0
0 100 200 300
Time (min)

pH 5

100

80

60

40
Release (%)

Simple
Composite
20 Coated 1 min
Coated 10 min

0
0 100 200 300
Time (min)

Figure 3. Cintique de libration de la riboflavine partir de 3 systmes (alginate seul,

composite alginate- gomme laque et enrobage alginate-gomme laque pendant 1 et 10
min par prcipitation acide la gomme diffrents pH ) [22].

24