U.S. patent application number 14/260384 was filed with the patent office on 2014-12-18 for adhering composition and methods of applying the same.
This patent application is currently assigned to TECHNION RESEARCH & DEVELOPMENT FOUNDATION LTD.. The applicant listed for this patent is TECHNION RESEARCH & DEVELOPMENT FOUNDATION LTD.. Invention is credited to Havazelet BIANCO-PELED, Ohad KIMHI.
Application Number | 20140370074 14/260384 |
Document ID | / |
Family ID | 40486892 |
Filed Date | 2014-12-18 |
United States Patent
Application |
20140370074 |
Kind Code |
A1 |
BIANCO-PELED; Havazelet ; et
al. |
December 18, 2014 |
ADHERING COMPOSITION AND METHODS OF APPLYING THE SAME
Abstract
A method of in-situ adhering comprising providing pre-gel that
comprises a mixture of at least one phenol-based compound and at
least one water miscible polymer selected from at least one of a
naturally existing form of a carbohydrate, a synthetically prepared
form of carbohydrate and a salt of an anionic polysaccharide;
spreading a layer of the pre-gel onto a first surface; adding a
solid support, comprising at least one cross linking agent capable
of interacting with the water miscible polymer to the pre-gel; and
allowing the pre-gel to cure and adhere onto the first surface.
Inventors: |
BIANCO-PELED; Havazelet;
(Haifa, IL) ; KIMHI; Ohad; (Kiryat Yam,
IL) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
TECHNION RESEARCH & DEVELOPMENT FOUNDATION LTD. |
Haifa |
|
IL |
|
|
Assignee: |
TECHNION RESEARCH & DEVELOPMENT
FOUNDATION LTD.
Haifa
IL
|
Family ID: |
40486892 |
Appl. No.: |
14/260384 |
Filed: |
April 24, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
12741495 |
Sep 13, 2010 |
8709480 |
|
|
PCT/IL2008/001451 |
Nov 5, 2008 |
|
|
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14260384 |
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61039564 |
Mar 26, 2008 |
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60985349 |
Nov 5, 2007 |
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Current U.S.
Class: |
424/443 ;
424/78.02; 424/78.18; 514/54 |
Current CPC
Class: |
A61L 15/585 20130101;
A61L 24/043 20130101; A61P 17/02 20180101; A61K 9/0024 20130101;
A61L 24/08 20130101 |
Class at
Publication: |
424/443 ; 514/54;
424/78.18; 424/78.02 |
International
Class: |
A61L 24/08 20060101
A61L024/08; A61L 24/04 20060101 A61L024/04 |
Claims
1. A method of in-situ adhering comprising: providing pre-gel that
comprises a mixture of at least one phenol-based compound and at
least one water miscible polymer selected from at least one of a
naturally existing form of a carbohydrate, a synthetically prepared
form of carbohydrate and a salt of an anionic polysaccharide;
spreading a layer of said pre-gel onto a first surface; adding a
solid support, comprising at least one cross linking agent capable
of interacting with said at least one water miscible polymer, to
said pre-gel; allowing said pre-gel to cure and adhere onto the
first surface.
2. A method of in-situ adhering comprising: providing pre-gel that
comprises at least one phenol-based compound, at least one water
miscible polymer selected from at least one of a naturally existing
form of a carbohydrate, a synthetically prepared form of
carbohydrate and a salt of an anionic polysaccharide, and a cross
linking agent comprising at least one water-insoluble salt of
multivalent ions capable of interacting with said at least one
water miscible polymer; spreading a layer of said pre-gel onto a
first surface; adding a solid support to said pre-gel; adding to
and blending with the pre-gel at least one trigger compound capable
of triggering release of multivalent ions from the cross linking
agent into the pre-gel, allowing the pre-gel to cure and adhere to
the first surface.
3. The method as claimed in claim 1, wherein said curing and
adhering is achieved by a method selected from one or more of a
group comprising spraying, dripping, and wetting the solid support
with said cross linking agent.
4. The method as claimed in claim 1, wherein said at least one
cross linking agent is provided within said solid support.
5. The method as claimed in claim 1, further comprising soaking
said solid support with said pre-gel.
6. The method as claimed in claim 1, further comprising embedding
said solid support within said pre-gel.
7. The method as claimed in claim 1, wherein said solid support is
coated with said cross linking agent.
8. The method as claimed in claim 1, further comprising drying said
pre-gel.
9. The method as claimed in claim 1, wherein said first surface is
a surface selected from a group of tissue surface, graft surface,
and organ surface.
10. The method as claimed in claim 2, wherein said first surface is
a surface selected from a group of tissue surface, graft surface,
and organ surface.
11. The method of adhering as claimed in claim 1, wherein said
adhering is under dry or wet conditions.
12. The method of claim 11, further comprising adhering the first
surface to a second surface, wherein each of said first surface and
said second surface is dry.
13. The method of claim 11, further comprising adhering the first
surface to a second surface, wherein at least one of said first
surface and said second surface is wet.
14. The method of claim 13, wherein at least one of said first
surface and said second surface is a body part or a component
thereof, of a human or animal subject.
15. The method of claim 14, wherein said component is a tissue.
16. The method of claim 1, wherein said adhering comprises sealing
or closing an opening in the first surface.
17. The method of claim 16, wherein said sealing or closing takes
place under dry or wet conditions.
18. The method of claim 16, wherein said surface having said
opening is a body part or a component thereof, of a human or animal
subject.
19. The method of claim 18, wherein said component is a tissue.
20. A method of treating an subject comprising: spreading a pre-gel
onto a tissue of the subject, the pre-gel comprising a mixture of
at least one phenol-based compound and at least one water miscible
polymer selected from at least one of a naturally existing form of
a carbohydrate, a synthetically prepared form of carbohydrate and a
salt of an anionic polysaccharide; adding a solid support,
comprising at least one cross linking agent capable of interacting
with said at least one water miscible polymer, to said pre-gel;
allowing said pre-gel to cure and adhere onto the tissue.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This patent application is a U.S. National Phase filing of
PCT patent application No. PCT/IL2008/001451, filed Nov. 5, 2008,
which is based upon U.S. provisional patent application No.
60/985,349, filed Nov. 5, 2007, and U.S. provisional patent
application No. 61/039,564, filed Mar. 26, 2008 all of which are
incorporated herein by reference. This application is a
continuation of patent application Ser. No. 12/741,495, now U.S.
Pat. No. 8,709,480.
FIELD OF THE INVENTION
[0002] The present invention relates to adhesives. More
particularly, the present invention relates to adhesive composition
of matter and methods of applying the same, especially in medical
applications.
BACKGROUND OF THE INVENTION
[0003] Tissue adhesives have been increasingly used to enhance
traditional closure technologies such as sutures and staples,
offering improved sealing capabilities and plugging of undesired
leaks'. However, despite recent developments and increased clinical
demand, currently available products still suffer from serious
drawbacks. While synthetic adhesives have low biocompatibility, low
adherence to wet surfaces and potential toxicity, the biological
glues are costly, often show relatively poor mechanical and
tissue-bonding properties, and are potentially immunogenic, as most
of them are based on proteins. Thus, there is a genuine unmet need
for non-toxic, strong, and economical tissue sealants to sustain
internal surgical incision closure, as an adjunct to suturing or
stapling. This need was the main motivation for the development of
bio-mimicking adhesives, which received increasing attention in the
last decade.
[0004] Using an adhesive for tissue reattachments or repair
procedures usually require the adhesive to be applied onto a
hydrated tissue surface. Moreover, biomedical adhesives have to
overcome contact with physiological fluids such as blood or saline
in order to form contacts or associations with the underlying
tissue. The success of synthetic adhesives in a hydrated
environment is limited, and typically requires certain treatments
and/or performing partial dehydration of the contact surface.sup.2.
In contrast to synthetic materials, nature has very effectively
conquered the limitations of sticking to wet surfaces.sup.3. Marine
sessile organisms such as barnacles, reef worms, mussels, algae
have life histories that depend on their secure attachment to solid
substrate for survival. These organisms produce and secrete
adhesives that form permanent, strong and flexible underwater bonds
to virtually any hard surface.sup.4. For example, mussels attach to
wet surfaces by creating a byssus, an extracorporeal bundle of tiny
tendons that are attached distally to a foreign substratum and
proximally by insertion of the stem root into the byssal retractor
muscles. "Mussel glues" have been proposed to be suitable for
medical applications due to their high adhesion strength and their
ability to adhere to wet surfaces. However, it is clear that the
commercial production of such glues is currently not practical,
since extraction of 1 kg of the naturally existing adhesive raw
materials (proteins and polypeptides) would require processing five
to ten million mussels.sup.1.
[0005] An alternative and more practical method is based on taking
a `biomimetic` approach, which entails constructing artificial
materials that mimic natural forms. Polymeric analogs may be
synthesized, as an example, from amino acids that were identified
as being functional to naturally existing adhesive proteins. Much
effort has been made to synthesize random block copolymers, which
are biomimetic approximations of naturally existing adhesive
proteins and polypeptides.sup.2,8-20.
[0006] Another effective natural adhesion mechanism exists in red
and brown algae, which produce phenolic compounds that exhibit
adhesive properties and extraordinarily high cohesive strength.
These adhesive contain phenolic compounds that bind
non-specifically to both hydrophobic and hydrophilic surfaces in
aqueous conditions.sup.21. The secretion of these phenolic
compounds is coupled with peroxidase oxidation and results in their
crosslinking of cell-wall polysaccharides. Based on those
observations, Vreeland et al. disclosed in U.S. Pat. No. 5,520,727
entitled "Aqueous algal-based phenolic type adhesives and glues" in
which the algal-based phloroglucinol was activated and cross linked
with algal carbohydrates in order to form glue. The inventors of
the present invention have demonstrated that formulations composed
of oxidized polyphenol extracted from Fucus serratus, alginate and
calcium ions are capable of adhering to a variety of
surfaces.sup.23. Structural analysis using small angle x-ray
scattering (SAXS) and electron microscopy (cryo-TEM) showed that
the polyphenols self-assemble into chain-like objects.sup.24.
Oxidation did not alter this overall structure, causing only a
reduction in the aggregate size. Moreover, this chain-like
structure did not change upon addition of alginate. Once calcium
ions were added, a network (whose overall structure resembled that
of the alginate gel) was formed.
[0007] Since the production of nature-based glues such as disclosed
in Vreeland et al. and others rely on extracting natural materials
from tons of algae, there was a need to synthetically imitate the
remarkable ability of marine algae to attach to wet solid surface
in order to provide effective adhesives having characteristics that
are similar to the characteristics of the marine algae.
[0008] Using the biomimetic approach, the inventors of the present
invention hypothesized that the natural components of the "fucus
glue" can be successfully replaced with commercially available
analogue that provides similar functionally. In PCT/IL2006/000289,
the inventors of the present invention indeed showed that the
monomeric unit of phloroglucinol and several of its derivatives to
interact with polysaccharide such as alginate to form an adhesive
that was shown to adhere in various compositions to animal tissues
as well as to other surfaces.
[0009] Interactions between carbohydrates and polyphenols are not
unique to algae adhesives. Polyphenols are a large and very diverse
family of plant metabolites, characterized by the presence of more
than one phenol group per molecule.sup.25-30.
[0010] It is needed to extend the adhesive composition beyond the
biomimetic approach and to develop adhesive composition of matter
that are able to form strong interactions with surfaces, whether
dry or wet, as well as within the network itself.
[0011] Moreover, the method of applying the adhesive materials
seems to play an important role in the ability to utilize the
adhesive composition of matter as an effective sealant. The
inventors of the present invention developed methods of applying
the adhesive material that allow on-site curing of the adhesive and
usage of bandages that form with the adhesive material an affective
sealant especially for medical use.
SUMMARY OF THE INVENTION
[0012] It is therefore provided in accordance with one aspect, a
method of in-situ adhering comprising: providing pre-gel that
comprises a mixture of at least one phenol-based compound and at
least one water miscible polymer selected from at least one of a
naturally existing form of a carbohydrate, a synthetically prepared
form of carbohydrate and a salt of an anionic polysaccharide;
spreading a layer of said pre-gel onto a first surface; adding a
solid support, comprising at least one cross linking agent capable
of interacting with said at least one water miscible polymer, to
said pre-gel; allowing said pre-gel to cure and adhere onto the
first surface.
[0013] It is therefore provided in accordance with another aspect,
a method of in-situ adhering comprising: providing pre-gel that
comprises at least one phenol-based compound, at least one water
miscible polymer selected from at least one of a naturally existing
form of a carbohydrate, a synthetically prepared form of
carbohydrate and a salt of an anionic polysaccharide, and a cross
linking agent comprising at least one water-insoluble salt of
multivalent ions capable of interacting with said at least one
water miscible polymer; spreading a layer of said pre-gel onto a
first surface; adding a solid support to said pre-gel; adding to
and blending with the pre-gel at least one trigger compound capable
of triggering release of multivalent ions from the cross linking
agent into the pre-gel, allowing the pre-gel to cure and adhere to
the first surface.
[0014] Optionally in accordance with another embodiment, said
curing and adhering is achieved by a method selected from one or
more of a group comprising spraying, dripping, and wetting the
solid support with said cross linking agent.
[0015] Optionally in accordance with another embodiment, said at
least one cross linking agent is provided within said solid
support.
[0016] Optionally in accordance with another embodiment, the method
further comprising soaking said solid support with said
pre-gel.
[0017] Optionally in accordance with another embodiment, the method
further comprising embedding said solid support within said
pre-gel.
[0018] Optionally in accordance with another embodiment, said solid
support is coated with said cross linking agent.
[0019] Optionally in accordance with another embodiment, the method
further comprising drying said pre-gel.
[0020] Optionally in accordance with another embodiment, said first
surface is a surface selected from a group of tissue surface, graft
surface, and organ surface.
[0021] Optionally in accordance with another embodiment, said first
surface is a surface selected from a group of tissue surface, graft
surface, and organ surface.
[0022] Optionally in accordance with another embodiment, said
adhering is under dry or wet conditions.
[0023] Optionally in accordance with another embodiment, the method
further comprising adhering the first surface to a second surface,
wherein each of said first surface and said second surface is
dry.
[0024] Optionally in accordance with another embodiment, the method
further comprising adhering the first surface to a second surface,
wherein at least one of said first surface and said second surface
is wet.
[0025] Optionally in accordance with another embodiment, at least
one of said first surface and said second surface is a body part or
a component thereof, of a human or animal subject.
[0026] Optionally in accordance with another embodiment, said
component is a tissue.
[0027] Optionally in accordance with another embodiment, said
adhering comprises sealing or closing an opening in the first
surface.
[0028] Optionally in accordance with another embodiment, said
sealing or closing takes place under dry or wet conditions.
[0029] Optionally in accordance with another embodiment, said
surface having said opening is a body part or a component thereof,
of a human or animal subject.
[0030] Optionally in accordance with another embodiment, said
component is a tissue.
[0031] It is also provided in accordance with yet another
embodiment, a method of treating an subject comprising: spreading a
pre-gel onto a tissue of the subject, the pre-gel comprising a
mixture of at least one phenol-based compound and at least one
water miscible polymer selected from at least one of a naturally
existing form of a carbohydrate, a synthetically prepared form of
carbohydrate and a salt of an anionic polysaccharide; adding a
solid support, comprising at least one cross linking agent capable
of interacting with said at least one water miscible polymer, to
said pre-gel; allowing said pre-gel to cure and adhere onto the
tissue.
BRIEF DESCRIPTION OF THE FIGURES
[0032] Some embodiments of the invention are herein described, by
way of example only, with reference to the accompanying drawings.
With specific reference now to the drawings in detail, it is
stressed that the particulars shown are by way of example and for
purposes of illustrative discussion of the preferred embodiments of
the present invention only, and are presented in the cause of
providing what is believed to be the most useful and readily
understood description of the principles and conceptual aspects of
the invention. In this regard, no attempt is made to show
structural details of the invention in more detail than is
necessary for a fundamental understanding of the invention, the
description taken with the drawings making apparent to those
skilled in the art how the several forms of the invention may be
embodied in practice.
[0033] In the drawings:
[0034] FIG. 1 illustrates several chemical structures of selected
phenols that can be utilized as adhesive base materials, in their
monomeric or polymeric forms.
[0035] FIG. 2 illustrates tensile strength required to separate two
porcine tissue strips adhered by polyphonel-based adhering
materials in accordance with preferred embodiments of the present
invention. The right bar represents the tensile strength obtained
with Tisseel.RTM., a commercial fibrin sealant.
[0036] FIG. 3 illustrates tensile strength of KI-oxidized adhesive
(dark grey) in accordance with a preferred embodiment of the
present invention compared to KI-oxidized Fucus glue (light grey)
to various substrates.
[0037] FIG. 4 illustrates SAXS curves from alginate and calcium ()
and phloroglucinol-containing adhesive material (.diamond.). The
solid lines represent the fit results of the "broken rod linked by
a flexible chain" model. Best-fit parameters are: Alginate gel:
.xi.=25 .ANG., R.sub.1=8.2 .ANG., R.sub.2=118 .ANG., C=8.1.
Biomimetic glue: .xi.=22 .ANG., R.sub.1=7.8 .ANG., R.sub.2=85
.ANG., C=6.7.
[0038] FIG. 5 illustrates ITC data for (A) Phloroglucinol (8 mg/ml,
63.4 mM) titrated into LF-200S alginate (5 mg/ml, 0.0236 mM). Lower
plot shows a fit to a two-site model. (B) Epicatechin (2 mg/ml,
68.8 mM) titrated into LF-200S alginate (5 mg/ml, 0.0236 mM). Lower
plot shows a fit to a one-site model.
DETAILED DESCRIPTION OF THE INVENTION AND THE FIGURES
[0039] The present invention provides novel composition-of-matter
of adhesives and methods of applying thereof in a wide variety of
different fields, and in particular, in the health care fields of
medicine, dentistry, and veterinary science. The present invention
is especially applicable for use by health care providers, such as
medical, dental, and veterinary, surgeons, in procedures for
reattaching or repairing body parts or components thereof, such as
tissues of human or animal subjects, especially under wet
conditions, but not necessarily. The composition-of-matter of the
present invention, applied as an adhesive, may also function, and
be usable, as a sealant or sealing agent, for sealing or closing an
opening in a surface, for example, for preventing flow of liquid
or/and gaseous fluid. Such a sealant or sealing agent can be used
in a wide variety of applications, for example, for sealing or
closing an opening in a medical device, of an aquarium, or of a
wide variety of other objects or entities.
[0040] A main aspect of the present invention is providing a
composition of matter that comprises at least one cross-linked form
of a water miscible polymer and at least one phenol-based material.
The phenol-based material can be in its monomeric or polymeric
form.
[0041] Accordingly and optionally, at least one activating agent is
used in the composition for promoting reaction and possible
cross-linking, or/and oxidation, or/and some other modification, of
any of the phenol type compound so as to produce its polymeric
form. Such activating agents are, for example, haloperoxidase (HPO)
enzyme, an oxidizer, a halogen salt, and combinations thereof.
[0042] The water miscible polymer is essentially any type or kind
of naturally existing polymer or synthetically prepared polymer
which is miscible in water. Examplary water miscible polymers can
be naturally existing, or synthetically prepared, form of a
carbohydrate (polysaccharide), such as alginic acid, or/and alginic
acid itself. More preferably, the water miscible polymer is a
naturally existing, or synthetically prepared, salt form of a
carbohydrate (polysaccharide), such as a salt form of alginic acid,
being an alginate. The water miscible polymer is in a cross-linked
form. For an embodiment of the composition-of-matter of the present
invention, wherein the water miscible polymer is an alginate, or
alginic acid, preferably, the alginate, or the alginic acid, is
cross-linked via interaction with divalent ions, for example,
divalent calcium ions (Ca.sup.+2) supplied, for example, by calcium
chloride (CaCl.sub.2), or by a combination of calcium carbonate
(CaCO.sub.3) and glucono-.delta.-lactone (GDL).
[0043] Another main aspect of the present invention is providing a
pre-gel that may be usable as an adhesive in combination with a
solid support. The combined pre-gel and solid support may be
functional and usable as a sealant or sealing agent, for sealing or
closing an opening in a surface so as to prevent as an example flow
of a fluid through the sealed or closed portion of the surface. The
sealing or closing may take place under dry or wet conditions.
[0044] Reference is made to FIG. 1 illustrating several chemical
structures of selected phenols that can be utilized as adhesive
base materials. The structures are shown in their monomeric form,
however, their polymeric form is being used also. These examples
are of non-toxic polyphenols can be utilized as base material for
an adhesive that can be used in the medical field. The exemplary
polyphenols shown in FIG. 1 are appropriate analogues that can
replace the natural components of glues such as "fucus glue" or
others that were mentioned in the background section of this
document. Most importantly, such commercially available polyphenols
are purchased from Sigma as well as other manufacturers. A large
selection of polyphenols was exploited in order to evaluate the
influence of their molecular parameters (molecular weight,
abundance of phenolic groups within the structure, molecular
flexibility) on the interactions with the carbohydrate on one hand,
and on the properties and the performance of the adhesive material,
on the other.
[0045] It should be noted that some polyphenols are only sparingly
soluble in water; however preliminary experience has shown that
occasionally the solubility in polysaccharide solutions is higher
compared to water.
[0046] One example for a polysaccharide that is included in the
preparation of the adhesive material of the present invention is a
commercial alginate supplied by FMC Biopolymer. Alginate is
structured as a block copolymer with blocks of .alpha.-L-gluronic
acid (G) and .beta.-D-mannuronic acid (M) alternating interrupted
by regions of more random distribution of M and G units. Sizes of
the three blocks can vary over a wide range, giving rise to
alginates of different properties. Two types of sodium alginate,
Protanal LF 200 S with G content of .about.70% and Protanal HF 120
RBS with G content of .about.50% were used. It should be noted
again that this alginate was used as an exemplary alginate and any
other miscible polymer can be used in order to prepare the adhesive
material.
[0047] Alternatively and optionally, alginate can be replaced with
a polysaccharide that can undergo gelation upon interaction with
ions and changes in temperature. In this case, the gels are dense
cross-linked polymeric matrices that can hinder solute diffusion.
Therefore drugs, growth factors, hormones, or any other therapeutic
agents, can be entrapped within the cured glue or sealant and will
be slowly released in the site of application.
[0048] In the specific case of alginate, gelation is induced by the
addition of multivalent ions. For example, calcium ions were added
either as CaCl.sub.2, or by blending the mixture with insoluble
salt such as CaCO.sub.3 or CaEGTA followed by addition of the
slowly hydrolyzing D-gluco-.delta.-lactone (GDL). Salts, which were
previously shown to alter alginate-polyphenol complexation.sup.31,
were added to some of the formulations. Optionally, other sources
of multivalent ions are polyelectrolytes, organic salts, and other
inorganic salts (e.g. Al, Ba).
[0049] As mentioned herein before, the method of applying the
adhesive material is one of the main aspects of the present
invention and special focus is given to combining solid support to
the adhesive material so as to form an adhering bandage. In the
following text, the term "pre-gel glue" refers to a composition of
matter that typically contains a polysaccharide such as alginate,
polyphenol, salt and water, at any given ratios. The pre-gel may
also contain multivalent ions, yet it has relatively low viscosity
since the polysaccharide is not fully gelled. The solid support may
be a thin film made from plastic (e.g. PGA, poly(caprolacton)
etc.), knitted mesh of fabric made from synthetic or natural
polymer, gauze prepared form oxidized cellulose etc. Any other
composition of the pre-gel as claimed in the present invention can
be used without limiting the scope of the present invention.
[0050] Optionally, the pre-gel glue may contain non-soluble
suspended solids in the form of particles, fibers etc. which are
added to enhance the mechanical strength of the glue.
[0051] The pre-gel glue is cured (solidified) so as to form an
adhesive material typically due to the addition of multivalent ions
or another method which induces polysaccharide gelation.
[0052] Several methods are used to apply the glue on site that
requires tissue repair or tissue sealing as follows:
[0053] (1) Spreading a layer of the pre-gel on the tissue or
surface followed by hardening (curing) of the polysaccharide which
can be achieved, for example, by spraying, dripping, or wetting the
pre-gel with an aqueous solution containing multivalent ions.
[0054] (2) Blending the pre-gel solution with insoluble salt of
multivalent ions (e.g. CaCO.sub.3 or CaEGTA) and an acid (e.g. the
slowly hydrolyzing D-gluco-.delta.-lactone (GDL)), then spreading a
layer on the tissue or the surface. The pre-gel hardens with time
due to the slow dissolution of the multivalent salt.
[0055] (3) Providing a patch for solid support such as a thin film
made from plastic (e.g. PGA, poly(caprolacton) etc.), knitted mesh
of fabric made from synthetic or natural polymer, gauze prepared
from oxidized cellulose etc. The patch or solid support is soaked
with a pre-gel and placed on the tissue. Alternatively, a layer of
pre-gel is spread on the tissue or the surface and the solid
support is embedded within it.
[0056] Finally, the polysaccharide is hardened by spraying,
dripping, or wetting the pre-gel with an aqueous solution
containing multivalent ions.
[0057] (4) Blending the pre-gel solution with insoluble salt of
multivalent ions (e.g. CaCO.sub.3 or CaEGTA) and an acid (e.g.
slowly hydrolyzing D-gluco-d-lactone (GDL) or acetic acid). A patch
or solid support such as a thin film made from plastic (e.g. PGA,
poly(caprolacton) etc.), knitted mesh of fabric made from synthetic
or natural polymer, gauze prepared from oxidized cellulose etc. is
soaked with a the pre-gel and placed on the tissue or the
surface.
[0058] Alternatively, a layer of pre-gel is spread on the tissue
and the solid support is embedded within it. The pre-gel hardens
with time due to the slow dissolution of the multivalent salt.
[0059] (5) Coating a solid support such as a thin film made from
plastic (e.g. PGA, poly(caprolacton) etc.), knitted mesh of fabric
made from synthetic or natural polymer, gauze prepared from
oxidized cellulose as an example with multivalent ions. A layer of
pre-gel is spread on the tissue or the surface and the solid
support is embedded in it. The pre-gel hardens with time due to the
slow release of the multivalent salt from the solid support.
[0060] (6) Preparing a dry film made from the components of the
pre-gel. Multivalent ions are used to coat a solid support as
described above, and the solid film is attached on top of the dried
pre-gel. Both films are placed on a hydrated tissue. The hydration
of the films leads to their adhering to the tissue. At the same
time, the pre-gel hardens with time due to the slow release of the
multivalent salt from the solid support.
[0061] (7) Preparing a dry film made from the components of the
pre-gel, with added multivalent ions in a dry form. The dry
multivalent ions may be incorporated in the pre-gel formulation or
as a different layer of the dry film (i.e. multilayer film). The
film is placed on a hydrated tissue or surface. The hydration of
the film leads to its adhering to the tissue. At the same time, the
pre-gel hardens with time due to the slow dissolution release of
the multivalent salt.
[0062] Regardless of the method by which the bandage was prepared,
additional top layers (typically made of polymers) may be added to
the finished bandage to provide extra mechanical support, provide
an inert layer that will separate between the inner bandage layers
and the surrounding environment etc. The additional layers, by no
means, limit the scope of the present invention.
[0063] In order to show the performance of the composition of
matter of the present invention as adhesives, several adhesive
properties were tested for several of the polyphenol-based adhesive
materials and the adhesive bandage.
Experimental Methods
Adhesive Properties
[0064] Adhesive strength is characterized using shear lap and
tensile tests. The samples holders and specimens preparation was
described in the inventor's PCT publication WO 06/092798. Briefly,
two identical sample holders are used to prepare a specimen. The
adhesive material is fixed to both sample holders using synthetic
glue. Next, a measured volume of the studied composition of matter
is applied onto one adherent, immediately covered with the other,
clamped, and cured for a specified time period. Adhesion tests are
performed using a Lloyd tensile machine equipped with a 50N load
cell. 10 specimens are prepared for each formulation and a
statistical analysis is performed. From practical reasons, and
following previous works, specimen preparation and adhesion assays
are performed at room temperature with no attempt to simulate
gluing under water..sup.2,8. However, evaluation of the influence
of curing in humid environment was established on the final
results.
[0065] The adhesion to different well-defined clean test substrates
(e.g. glass, metals, and different types of plastic) is
characterized for the different composition of matters.
[0066] Additional adhesion assays were performed using tissues
obtained from a local slaughterhouse. These provide better sense
for the glue's ability to adhere to moist and flexible
surfaces.
Rheological Measurements
[0067] Rheological measurements were conducted using a Rheometric
Scientific ARES strain-controlled rheometer fitted with a 50-mm
cone-and-plate fixture and equipped with anti-evaporator cover that
prevents sample dehydration. Oscillatory shear experiments were
performed within the linear viscoelastic regime, where the dynamic
storage modulus (G') and loss modulus (G'') are independent of the
strain amplitude. Calculations are done using the RSI Orchestrator
6.5.1 software. The critical calcium concentration required to
induced a sol-gel transition at a given composition of matter
concentration are determined from the appearance of a power law of
the dynamic moduli.sup.35, as proposed by Winter and
Chambon.sup.36,37. Moduli exhibiting power law dependency on the
frequency .omega. of G'.about..omega..sup.1.5 and
G''.about..omega..sup.1 are characteristic to a viscoelastic fluid
according to the Rouse-Zimm theory. Above the gel point, G' becomes
larger than G'' with a plateau appearing in the G' vs. .omega.
curve at the low frequency range.
Wetting Properties
[0068] Advancing contact angles of the liquids on the test surfaces
are measured by a computerized contact angle analyzer (CAM200, KSV,
Helsinki, Finland). A micro Hamilton syringe were used to create
the drop on the surface and to continuously add liquid during the
measurement. The surface tension of the aqueous solutions was
measured by the ring method. The work of the adhesion W.sub.adh is
calculated according to the Young-Dupre equation
W.sub.adh=.gamma..sub.LV (1 .theta.).sup.8. The wetting properties
of both polyphenol solution and polyphenol/alginate was
studied.
Miscible Polymer/Polyphenol Interactions
[0069] .sup.1H NMR Spectroscopy: Comparing spectra obtained from
the composition of matter solutions with spectra obtained from the
individual components provide a qualitative indication to
intramolecular interactions.sup.24.
[0070] Equilibrium Dialysis was previously used to determine the
extent of binding of polyphenols to macromolecules.sup.26. Binding
assays include dialyzing 0.5 ml polyphenol solution, contained in a
Spectra/Por.RTM. dialysis tubing (Spectrum Laboratories.RTM., CA,
USA) against stirred polysaccharide solution.sup.38. Once
equilibrium is achieved, the polyphenol concentration in both cells
is determined spectrophotometrically. The partition coefficient is
calculated from a mass balance. A control experiment (dialysis
against water) is used to ensure that polyphenol is not adsorbed by
the cellulose membrane installed in the dialysis tube.
[0071] Isothermal Titration calorimetry (ITC) is used to quantify
the magnitude of the interactions between the polyphenols and the
polysaccharides (alginate). ITC is a technique that allows studying
the heat of interaction between two molecules. Often, a "ligand" is
titrated to a solution of a "macromolecule", and the heat released
upon their interaction, Q, is monitored over time. As the two
elements interact, heat is released or absorbed in direct
proportion to the amount of association/dissociation that occurs.
When the ligand in the cell becomes saturated with added
macromolecule, the heat signal diminishes until only the background
heat of dilution is observed. ITC measurements were performed using
a VP-ITC Microcalorimeter (MicroCal Inc.). ITC experiments include
injecting known amounts of polyphenol solution into an aqueous
alginate solution. Appropriate reference experiments (injection of
buffer into alginate and polyphenol dilution) are performed as
well. The experimental data were analyzed using the software
provided with the instrument, which calculates the binding constant
from the slope of the heat vs. injected amount at the saturation
point. In a case of small binding energies, one of the analysis
scheme previously applied by the inventors of the present invention
and others.sup.39-41 was followed. Data obtained from ITC and
equilibrium binding experiments can be further used to calculate
the free energy and entropy of binding.sup.26,40. This approach
provides additional insight into the binding mechanism and in
particular highlights the related importance of the entropy driven
hydrophobic interactions and the enthalpy driven hydrogen
bonding.
Nanostructure Characterization
[0072] Nanostructure characterization allows assessment of the
estimated model of polyphenols and alginate that is hypnotized to
form a nanocomposite material. Moreover, the structural parameters
of such nanocomposite might have a significant impact on its
properties.
[0073] SAXS measurements are mostly performed using a
slit-collimated compact Kratky camera (A. Paar Co.) equipped with a
linear position sensitive detector system (Raytech), with
pulse-height discrimination and a multichannel analyzer (Nucleus).
A total of 3000 or more counts for each channel are collected in
order to obtain a high signal to noise ratio. Primary beam
intensities is determined using the moving slit method.sup.42 and
subsequently using a thin quartz monitor as a secondary standard.
The scattering curves, as a function of the scattering vector
q=4.pi. sin .theta./.lamda. (where 2.theta. and .lamda. are the
scattering angle and the wavelength, respectively), are corrected
for counting time and for sample absorption and the background is
subtracted. Desmearing procedure is performed according to the
Indirect Transformation Method.sup.43 using the ITP program. Data
analysis includes fitting the desmeared curve to an appropriate
model using a least-squares procedure. As a starting point, the
inventors fitted the model used for the algal-born glue.sup.24,
known as "broken rod linked by a flexible chain"
model.sup.44,45:
I ( q ) .varies. S ( q ) P ( q ) .varies. 1 1 + C exp ( - .xi. 2 q
2 ) s ( q ) 1 q 2 i k i q [ J 1 ( qR i ) qR i ] 2 P ( q )
##EQU00001##
[0074] Where P(q) is the form factor of cylindrical elements
specified by cross-sectional radii R.sub.i, and relative weights of
k.sub.i. The electrostatic interactions are taken into account by
the structure factor S(q) that assumes a Gaussian-type interaction
potential specified by the correlation length C is a an adjustable
parameter representing the strength of interaction, which depends
on the second virial coefficient and the polymer
concentration..sup.44 45.
[0075] Cryo-Transmission Electron Microscopy (cryo-TEM) is used in
parallel for direct visualization of the nanostructure. Ultra-fast
cooled vitrified specimens, prepared at controlled conditions of
25.degree. C. and 100% relative humidity as described
elsewhere.sup.46, are studied in a Philips CM120 cryo-TEM operating
at 120 kV. An Oxford CT3500 cooling-holder system that keeps the
specimens at about -180.degree. C. was used. Low electron-dose
imaging is performed with a Gatan Multiscan 791 CCD camera, using
the Gatan Digital Micrograph 3.1 software package.
Pulsatile Flow System.
[0076] A flow system was built in order to serve as a model system
that imitates the forces working on blood vessels in the human
body, thus providing a more realistic method of testing the
adhesives. The flow system creates pressure swing between the
systolic and diastolic blood pressures. The inventors tested
punctured aorta originated from bovine or swine as a substrate for
testing.
[0077] In order to analyze the results, an experimental parameter
named sealing ratio (SR) was defined as
SR = 1 Flow through sealed hole Flow rate through unsealed hole .
##EQU00002##
T-Peel Test
[0078] T-peel test is based on ASTM F-2256-05 that was developed as
testing method for surgical adhesives and sealants.
[0079] The samples used for the T-peel test are of constant
dimensions.
[0080] The solid support dimensions are 150.+-.1 mm length and
25.+-.1 mm width. The dimensions of the sample tissue, which was an
artery, equal to those of the solid support.
[0081] 1.75 ml of adhesive was applied on the tissue. The hardening
film is then applied and a weight of 1 kg is placed over it for one
minute. The sample is then placed in the extension test machine and
the test is conducted.
[0082] The results are analyzed as described in ASTM F-2256-05.
Experimental Results
[0083] Reference is now made to FIG. 2 illustrating tensile
strength required to separate two porcine tissue strips adhered by
polyphonel-based adhering materials in accordance with preferred
embodiments of the present invention. The right bar represents the
tensile strength obtained with Tisseel.RTM., a commercial fibrin
sealant, for comparison. Results show firm adhesion between the
tissue surfaces after adherence of the tissues with a phenolic
adhering material; wherein the tissues are porcine muscle
tissues.
[0084] Different types of polyphenols were used to demonstrate the
feasibility of the present invention. The composition of the
different polyphenol-based adhesive materials shows significant
adhesion properties. It should be noted that some of the
polyphenol-based adhesive materials showed better adhesion to the
tissue than Tisseel.RTM., a commercial fibrin sealant.
[0085] Reference is now made to FIG. 3 illustrating tensile
strength of KI-oxidized adhesive (dark grey) in accordance with a
preferred embodiment of the present invention compared to
KI-oxidized Fucus glue (light grey) to various substrates. Tensile
strength of alga-born glue, composed of components extracted from
Fucus Serratus (15 mg/ml Alginate, 5 mg/ml polyphenol, 4 mM ca
ions) are compared to these of a biomimetic composition in matter
in which the algae polyphenol was replaced with phloroglucinol and
the algae alginate with Protanal LF 200 S alginate. All samples
were oxidized by adding 0.75 U/ml BPO, 0.44% H.sub.2O.sub.2 and 4.4
mg/ml KI. Oxidation was verified using NMR (data not shown).
Samples were cured for 20 minutes. The adhesive strengths of the
biomimetic adhesive was comparable to that of the alga-born one.
Moreover, both glues seem to adhere better to the more hydrophobic
surfaces.
[0086] A preliminary examination of the effect of oxidation was
achieved by repeating the experiment described in (A) while
omitting the oxidation agents. Oxidized formula adhered better to
the Teflon while the non-oxidized formula adhered better to the
glass and Mylar (not shown).
[0087] Reference is now made to FIG. 4 illustrating SAXS curves
from alginate (15 mg/ml) and calcium (4 mM) polyphenol, 0.75 U/ml
BPO, 0.44% H.sub.2O.sub.2, 4.4 mg/ml KI, 15 mg/ml alginate and 4 mM
calcium ions was () and phloroglucinol-containing adhesive material
(alginate 15 mg/ml, phlologlucinol 5 mg/ml, Ca 4 mM) (.diamond.).
The solid lines represent the fit results of the "broken rod linked
by a flexible chain" model. Best-fit parameters are: Alginate gel:
.xi.=25 .ANG., R.sub.1=8.2 .ANG., R.sub.2=118 .ANG., C=8.1.
Biomimetic glue: .xi.=22 .ANG., R.sub.1=7.8 .ANG., R.sub.2=85
.ANG., C=6.7. The SAXS results comparison demonstrates the
existence of alginate-phloroglucinol interactions in a biomimetic
composition of matter. The overall structure of the glue resembles
that of alginate gel. Yet, since the scattering from phloroglucinol
by itself was very weak and a scattering curve could not be
obtained, the differences between the two curves can be attributed
to interactions between the alginate and the phloroglucinol. As in
the case of algal-born glue.sup.24, both curves were well fitted by
the "broken rod linked by a flexible chain" model. The analysis of
the SAXS data showed that the two samples differ by the correlation
length .xi., the adjustable parameter C and R.sub.2 that represents
the alginates' tendency to form large aggregates.sup.47. Smaller
correlation length and adjustable parameter were obtained for the
biomimetic glue, thus indicating weaker electrostatic repulsion and
lower hydrophilicity compared to alginate gel.sup.48.
[0088] Reference is now made to FIG. 5 illustrating ITC data for
(A) Phloroglucinol (8 mg/ml, 63.4 mM) titrated into LF-2005
alginate (5 mg/ml, 0.0236 mM). Lower plot shows a fit to a two-site
model. (B) Epicatechin (2 mg/ml, 68.8 mM) titrated into LF-200S
alginate (5 mg/ml, 0.0236 mM). Lower plot shows a fit to a one-site
model. Two preliminary ITC measurements detected heat changes
during injection of polyphenol into alginate solution, thus
providing additional evidence to interactions between the two
materials that comprises the adhesive material. Moreover, the
affect of the polyphenol's molecular structure is evident from a
comparison between the experiments.
[0089] As mentioned herein above, many different configurations of
dry film may be used in order to apply the adhering material of the
present invention with their formulations. The composition of
matter of the adhering material can differ in their physical form,
composition and preparation method. The following examples include
two different preparation methods and two different
compositions.
[0090] Air drier film, a solution containing 35 mg/ml
alginate(LF200S), 10 mg/ml PHG and 1 mg/ml colorant was cast into
molds with different depths and air dried for 24 hrs. The film was
peeled and use in conjunction with ORC containing .about.1
mg/cm.sup.2 CaCl2 as hardener. The sealing ratio was measured using
the flow system mentioned above. A sealing ratio close to 1 at 120
mmHg was achieved for 14 samples.
[0091] Freeze dried film are prepared from a solution containing
alginate, PHG and colorant that were cast into molds with different
widths. The solutions were freeze by liquid nitrogen, however, can
be freeze also by solid carbon dioxide or any other cooling method.
In some cases, a second layer was applied, wherein this layer
contained PVA/PEG/CACl.sub.2 mixture. The samples were than
inserted into freeze drier until completely dried.
[0092] Single layer samples--these patches were used in conjunction
with external ORC hardening system containing .about.1 mg/cm2
CaCl.sub.2. The solution used for casting these films contained 35
mg/ml alginate (LF200S), 10 mg/ml PHG and 0.3 mg/ml colorant (acid
green 25 or other such as indigo carmine)
[0093] The sealing ratio for 1.5 mm thick film was approximately 1
at 200 mmHg, The number of repetitions was 22.
[0094] Multilayer samples--a first 1.5 mm thick layer was cast
using solution containing 35 mg/ml alginate (LF200S), 10 mg/ml PHG
and 0.3 mg/ml colorant (acid green 25 or other such as indigo
carmine). After the first layer was frozen, a second layer was cast
using a solution containing PVA/PEG/CaCl2 in the following
concentrations, 34.4/15.6/20 mg/ml. The second layer was allowed to
freeze and the multilayer film was placed on freeze drier until
completely dried. The sealing ratio for this composition was found
to be approx. 1 at 200 mmHg.
[0095] Although most of the results shown herein deal with alginate
as a preferred water miscible polymer, a carbohydrate, other water
miscible polymers were tested. As an example, a T-peel test was
conducted for polygalacturonic acid as another preferred
carbohydrate that shows T-peel strength of 0.076N/cm.
[0096] For any of the above described preferred embodiments or
formulations of the composition of matter of the present invention
generally usable as an adhesive; the adhesive may be functional and
usable as sealant or sealing agent, for sealing or closing an
opening of a dry or wet surfaces. For example, for preventing flow
of a liquid or/and gaseous fluid through the sealed or closed
portion of the surface. Accordingly, the sealing or closing may
take place under dry or wet conditions. The surface having the
opening which is sealed or closed may be a body part or a component
thereof (e.g. a tissue), of a human or animal subject. Such a
sealant or sealing agent can be used in a wide variety of
applications, for example, for sealing or closing an opening in a
dry or wet body part, or in a dry or wet surface of a medical
device, of an aquarium, or of a wide variety of other objects or
entities.
[0097] Any of the above described preferred embodiments or
formulations of the composition of matter of the present invention
are generally usable as an adhesive, of a variety of different
types of surfaces, under dry or wet conditions.
[0098] In particular, the composition of matter of the present
invention is generally usable as an adhesive under dry conditions,
for example, for adhering a first surface to a second surface,
wherein both surfaces are dry. Alternatively and advantageously,
the composition of matter of the present invention is generally
usable as an adhesive under wet conditions, for example, for
adhering a first surface to a second surface wherein the first
surface is wet and/or the second surface is wet.
[0099] Any of the above described preferred embodiments or
formulations of the composition of matter of the present invention
is generally usable as a sealant or an adhesive in any condition,
wet or dry, and on any kind of surface, in a wide variety of
different fields such as health care, medicine, dentistry,
veterinary, as well as other general fields such as commercial,
laboratory or home use.
[0100] It should be clear that the description of the
configurations and attached Figures set forth in this specification
serves only for a better understanding of the invention, without
limiting its scope as covered by the following Claims.
[0101] It should also be clear that after reading the present
specification a skilled person, can make adjustments or amendments
to the attached Figures and above described configurations that
would still be covered by the following Claims.
REFERENCES
[0102] (1) Webster, I.; West, P. J. "Adhesives for medical
applications" In Polymeric Biomaterials, Dumitriu, S., Eds.; Marcel
Dekker, Inc.: New York, 2002; [0103] (2) Yu, M.; Deming, T. J.
"Synthetic Polypeptide Mimics of Marine Adhesives" Macromolecules
1998, 31, 4739-4745. [0104] (3) Callow, M., E.; Callow, J., E.
"Marine biofouling: a sticky problem" Biologist (London, England)
2002, 49, 10-14. [0105] (4) Hwang, D. S.; Yoo, H. J.; Jun, J. H.;
Moon, W. K.; Cha, H. J. "Expression of functional recombinant
mussel adhesive protein Mgfp-5 in Escherichia coli" Appl. Environ.
Microbiol. 2004, 70, 3352-3359. [0106] (5) Waite, J. H. "Nature's
underwater adhesive specialist" International Journal of Adhesion
and Adhesives 1987, 7, 9-14. [0107] (6) Waite, J. H. "Reverse
engineering of bioadhesion in marine mussels" Ann. N.Y. Acad. Sci.
1999, 875, 301-309. [0108] (7) Vreeland, V.; Waite, J. H.; Epstein,
L. "Polyphenols and oxidases in substratum adhesion by marine algae
and mussels" Journal of Phycology 1998, 34, 1-8. [0109] (8)
Yamamoto, H.; Sakai, Y.; Ohkawa, K. "Synthesis and wettability
characteristics of model adhesive protein sequences inspired by a
marine mussel" Biomacromolecules 2000, 1, 543-551. [0110] (9)
Ohkawa, K.; Ichimiya, K.; Nishida, A.; Yamamoto, H. "Synthesis and
surface chemical properties of adhesive protein of the asian
freshwater mussel, Limnoperna fortunei" Macromolecular Bioscience
2001, 1, 376-386. [0111] (10) Tatehata, H.; Mochizuki, A.;
Kawashima, T.; Yamashita, S.; Ohkawa, K.; Yamamoto, H. "Oxidative
reaction and conformational studies on synthetic sequential
polypeptides of mussel adhesive proteins" Current Trends in Polymer
Science 2000, 5, 91-96. [0112] (11) Tatehata, H.; Mochizuki, A.;
Kawashima, T.; Yamashita, S.; Yamamoto, H. "Model polypeptide of
mussel adhesive protein. I. Synthesis and adhesive studies of
sequential polypeptides (X-Tyr-Lys).sub.n and (Y-Lys).sub.n" J.
Appl. Polym. Sci. 2000, 76, 929-937. [0113] (12) Yamamoto, H.;
Nishida, A.; Ohkawa, K. "Wettability and adhesion of marine and
related adhesive proteins" Colloids and Surfaces, A:
Physicochemical and Engineering Aspects 1999, 149, 553-559. [0114]
(13) Yamamoto, H.; Ogawa, T.; Nishida, A. "Molecular weight
dependence of wettability and molecular adsorption of poly-L-lysine
at the air-water interface" J. Colloid Interface Sci. 1995, 176,
105-110. [0115] (14) Wang, J.; Liu, C.; Lu, X.; Yin, M.
"Co-polypeptides of 3,4-dihydroxyphenylalanine and -lysine to mimic
marine adhesive protein" Biomaterials 2007, 28, 3456-3468. [0116]
(15) Lee, B. P.; Dalsin, J. L.; Messersmith, P. B. "Synthesis and
Gelation of DOPA-Modified Poly(ethylene glycol) Hydrogels"
Biomacromolecules 2002, 3, 1038-1047. [0117] (16) Lee, B. P.;
Huang, K.; Nunalee, F. N.; Shull, K. R.; Messersmith, P. B.
"Synthesis of 3,4-dihydroxyphenylalanine (DOPA) containing monomers
and their co-polymerization with PEG-diacrylate to form hydrogels"
J. Biomater. Sci. Polym. Ed. 2004, 15, 449-464. [0118] (17) Catron,
N. D.; Lee, H.; Messersmith, P. B. "Enhancement of poly(ethylene
glycol) mucoadsorption by biomimetic end group functionalization"
Biointerphases 2006, 1, 134-141. [0119] (18) Messersmith, P. B.
"Mussel adhesive protein mimetics: polymer-peptide bioconjugates
for tissue adhesion and antifouling surfaces" PMSE Preprints 2006,
94, 129. [0120] (19) Huang, K.; Lee, B.; Messersmith, P. B.
"Synthesis and characterization of self-assembling block copolymers
containing adhesive moieties" Polym. Prepr. (Am. Chem. Soc., Div.
Polym. Chem.) 2001, 42, 147-148. [0121] (20) Westwood, G.; Horton,
T. N.; Wilker, J. J. "Simplified Polymer Mimics of Cross-Linking
Adhesive Proteins" Macromolecules (Washington, D.C., U.S.) 2007,
40, 3960-3964. [0122] (21) Vreeland, V.; Epstein, L. "Analysis of
plant-substratum adhesives" Modern Methods of Plant Analysis 1996,
17, 95-116. [0123] (22) Berglin, M.; Delage, L.; Potin, P.; Vilter,
H.; Elwing, H. "Enzymatic Cross-Linking of a Phenolic Polymer
Extracted from the Marine Alga Fucus serratus" Biomacromolecules
2004, 5, 2376-2383. [0124] (23) Bitton, R. "The Influence of
Halide-Mediated Oxidation on Algae-Born Adhesives" Macromolecular
Bioscience 2007, In press, [0125] (24) Bitton, R.; Ben-Yehuda, M.;
Davidovich, M.; Balazs, Y.; Potin, P.; Delage, L.; Colin, C.;
Bianco-Peled, H. "Structure of algal-born phenolic polymeric
adhesives" Macromolecular Bioscience 2006, 6, 737-746. [0126] (25)
Cai, Y.; Gaffney, S. H.; Lilley, T. H.; Magnolato, D.; Martin, R.;
Spencer, C. M.; Haslam, E. "Polyphenol interactions. Part 4. Model
studies with caffeine and cyclodextrins" Journal of the Chemical
Society, Perkin Transactions 2: Physical Organic Chemistry
(1972-1999) 1990, 2197-2209. [0127] (26) McManus, J. P.; Davis, K.
G.; Beart, J. E.; Gaffney, S. H.; Lilley, T. H.; Haslam, E.
"Polyphenol interactions. Part 1. Introduction: some observations
on the reversible complexation of polyphenols with proteins and
polysaccharides" Journal of the Chemical Society, Perkin
Transactions 2: Physical Organic Chemistry (1972-1999) 1985,
1429-1438. [0128] (27) Gaffney, S. H.; Martin, R.; Lilley, T. H.;
Haslam, E.; Magnolato, D. "The association of polyphenols with
caffeine and a- and b-cyclodextrin in aqueous media" J. Chem. Soc.,
Chem. Commun. 1986, 107-109. [0129] (28) Renard, C. M. G. C.;
Baron, A.; Guyot, S.; Drilleau, J. F. "Interactions between apple
cell walls and native apple polyphenols: quantification and some
consequences" Int. J. Biol. Macromol. 2001, 29, 115-125. [0130]
(29) Bourvellec, C. L.; Bouchet, B.; Renard, C. M. G. C.
"Non-covalent interaction between procyanidins and apple cell wall
material. Part III: Study on model polysaccharides" Biochim.
Biophys. Acta 2005, 1725, 10-18. [0131] (30) Nishinari, K.; Kim,
B.; Fang, Y.; Nitta, Y.; Takemasa, M. "Rheological and related
study of gelation of xyloglucan in the presence of small molecules
and other polysaccharides" Cellulose (Dordrecht, Netherlands) 2006,
13, 365-374. [0132] (31) Haslam, E.; Lilley, T. H.; Warminski, E.;
Liao, H.; Cai, Y.; Martin, R.; Gaffney, S. H.; Goulding, P. N.;
Luck, G. "Polyphenol complexation. A study in molecular
recognition" In Phenolic Compounds in Foods and Their Effects on
Health I, Analysis, Occurrence & Chemistry, Ho, C.-T.; Lee, C.
Y.; Huang, M.-T., Eds.; American Chemical Society Washington, D.C.,
1992; 506. [0133] (32) Sklenar, V.; Piotto, M.; Leppik, R.; Saudek,
V. "Gradient-tailored water suppression for proton-nitrogen-15 HSQC
experiments optimized to retain full sensitivity" Journal of
Magnetic Resonance, Series A 1993, 102, 241-245. [0134] (33)
Piotto, M.; Saudek, V.; Sklenar, V. "Gradient-tailored excitation
for single-quantum NMR spectroscopy of aqueous solutions" J.
Biomol. NMR 1992, 2, 661-665. [0135] (34) Manderville, R. A.
"Ambident reactivity of phenoxyl radicals in DNA adduction" Can. J.
Chem. 2005, 83, 1261-1267. [0136] (35) Liu, X.; Qian, L.; Shu, T.;
Tong, Z. "Rheology characterization of sol-gel transition in
aqueous alginate solutions induced by calcium cations through in
situ release" Polymer 2002, 44, 407-412. [0137] (36) Chambon, F.;
Winter, H. H. "Linear viscoelasticity at the gel point of a
crosslinking PDMS with imbalanced stoichiometry" Journal of
Rheology (New York, N.Y., U.S.) 1987, 31, 683-697. [0138] (37)
Winter, H. H.; Chambon, F. "Analysis of linear viscoelasticity of a
crosslinking polymer at the gel point" Journal of Rheology (New
York, N.Y., U.S.) 1986, 30, 367-382. [0139] (38) Regev, R.;
Yeheskely-Hayon, D.; Katzir, H.; Eytan, G. D. "Transport of
anthracyclines and mitoxantrone across membranes by a flip-flop
mechanism" Biochem. Pharmacol. 2005, 70, 161-169. [0140] (39)
Kimhi, 0.; Bianco-Peled, H. "Study of the Interactions between
Protein-Imprinted Hydrogels and Their Templates" Langmuir 2007, 23,
6329-6335. [0141] (40) Bianco-Peled, H.; Gryc, S. "Binding of Amino
Acids to \"Smart\" Sorbents: Where Does Hydrophobicity Come into
Play?" Langmuir 2004, 20, 169-174. [0142] (41) Kimhi, 0.;
Bianco-Peled, H. "Microcalorimetry Study of the Interactions
between Poly(N-isopropylacrylamide) Microgels and Amino Acids"
Langmuir 2002, 18, 8587-8592. [0143] (42) Stabinger, H.; Kratky, 0.
"A new technique for the measurement of the absolute intensity of
x-ray small angle scattering. The moving slit method" Makromol.
Chem. 1978, 179, 1655-1659. [0144] (43) Glatter, 0. "A New Method
for the Evaluation of Small-Angle Scattering Data" j.Appl.Cryst
1977, 10, 415-421. [0145] (44) Yuguchi, Y.; Urakawa, H.; Kajiwara,
K.; Draget, K. I.; Stokke, B. T. "Small-angle x-ray scattering and
rheological characterization of alginate gels. 2. Time-resolved
studies on ionotropic gels" J. Mol. Struct. 2000, 554, 21-34.
[0146] (45) Yuguchi, Y.; Mimura, M.; Urakawa, H.; Kitamura, S.;
Ohno, S.; Kajiwara, K. "Small angle x-ray characterization of
gellan gum containing a high content of sodium in aqueous solution"
Carbohydr. Polym. 1996, 30, 83-93. [0147] (46) Bellare, J. R.;
Davis, H. T.; Scriven, L. E.; Talmon, Y. "Controlled environment
vitrification system: an improved sample preparation technique" J.
Electron Microsc. Tech. 1988, 10, 87-111. [0148] (47) Windhues, T.;
Borchard, W. "Effect of acetylation on physico-chemical properties
of bacterial and algal alginates in physiological sodium chloride
solutions investigated with light scattering techniques" Carbohydr.
Polym. 2003, 52, 47-52. [0149] (48) Shimode, M.; Mimura, M.;
Urakawa, H.; Yamanaka, S.; Kajiwara, K. "Dye aggregation observed
by small-angle x-ray scatterings. Part 2. Interaction between the
dye aggregates in aqueous solution" Sen'l Gakkaishi 1996, 52,
301-309.
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