U.S. patent application number 13/561995 was filed with the patent office on 2013-03-28 for intrauterine device, method of making such a device and method for putting active elements within the uterine cavity.
This patent application is currently assigned to ANECOVA SA. The applicant listed for this patent is Pascal Mock. Invention is credited to Pascal Mock.
Application Number | 20130074846 13/561995 |
Document ID | / |
Family ID | 23197486 |
Filed Date | 2013-03-28 |
United States Patent
Application |
20130074846 |
Kind Code |
A1 |
Mock; Pascal |
March 28, 2013 |
INTRAUTERINE DEVICE, METHOD OF MAKING SUCH A DEVICE AND METHOD FOR
PUTTING ACTIVE ELEMENTS WITHIN THE UTERINE CAVITY
Abstract
A retrievable intrauterine device for placing within the uterine
cavity one or more encapsulated elements (1) capable of having
interactions with the uterine fluid comprising an intrauterine
device loaded with said encapsulated elements (1).
Inventors: |
Mock; Pascal; (Geneve,
CH) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Mock; Pascal |
Geneve |
|
CH |
|
|
Assignee: |
ANECOVA SA
Geneve
CH
|
Family ID: |
23197486 |
Appl. No.: |
13/561995 |
Filed: |
July 30, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10485611 |
Aug 18, 2004 |
8257244 |
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PCT/IB02/03363 |
Jul 22, 2002 |
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13561995 |
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60309274 |
Aug 1, 2001 |
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Current U.S.
Class: |
128/833 |
Current CPC
Class: |
A61F 6/144 20130101;
A61F 6/18 20130101; A61B 17/435 20130101 |
Class at
Publication: |
128/833 |
International
Class: |
A61F 6/18 20060101
A61F006/18 |
Claims
1-15. (canceled)
16. A retrievable intrauterine device comprising at least one
non-biodegradable capsule, wherein said capsule is adapted to
encapsulate one or more element(s) comprising an egg, an embryo,
male gametes, female gametes, a fertilized oocyte, a zygote, or a
combination thereof, further wherein said capsule comprises a
membrane with a pore size adapted to allow passage of nutrients
from a uterine cavity to the elements when the elements are loaded
in said capsule.
17. The retrievable intrauterine device of claim 16, wherein the
nutrients comprise proteins.
18. The retrievable intrauterine device of claim 16, further
comprising a means for retrieving the device from a uterus to
collect said elements(s).
19. The retrievable intrauterine device of claim 16, further
comprising a thread to retrieve the device from the uterus to
collect the elements(s).
20. The retrievable intrauterine device of claim 16, wherein said
capsule comprises a wall having a thickness ranging from 50 to 500
.mu.m.
21. The retrievable intrauterine device of claim 16, wherein the
length of said device ranges from 0.5 cm to 5 cm.
22. The retrievable intrauterine device of claim 16, wherein said
device comprises means for attachment of the device to the uterine
cavity.
23. The retrievable intrauterine device of claim 16, wherein said
capsule has an internal diameter ranging from 100 to 10,000
.mu.m.
24. The retrievable intrauterine device of claim 16, wherein said
capsule comprises a polymeric material.
25. A retrievable intrauterine device comprising at least one
non-biodegradable capsule, wherein said capsule comprises one or
more element(s) comprising an egg, an embryo, male gametes, female
gametes, a fertilized oocyte, a zygote, or a combination thereof,
further wherein said capsule comprises a means for enabling said
elements to interact with a uterine fluid.
26. The retrievable intrauterine device of claim 25, wherein the
means for enabling said elements to interact with a uterine fluid
comprises a membrane.
27. The retrievable intrauterine device of claim 25, wherein said
elements comprise male gametes or female gametes.
28. The retrievable intrauterine device of claim 25, wherein said
element comprises an embryo.
29. The retrievable intrauterine device of claim 25, wherein said
elements comprise injected oocytes.
30. The retrievable intrauterine device of claim 25, wherein said
elements comprise impregnated oocytes.
31. A retrievable intrauterine device comprising at least one
non-biodegradable capsule, wherein said capsule is adapted to
encapsulate one or more element(s) comprising an egg, an embryo,
male gametes, female gametes, a fertilized oocyte, a zygote, or any
combination thereof, further wherein said capsule comprises a means
for enabling said encapsulated elements to interact with a uterine
fluid, when the elements are encapsulated in said capsule.
Description
[0001] The present invention relates generally to intrauterine
devices and to method for placing elements into the uterine cavity
notably for therapeutic purposes. The uterine cavity is an
anatomical location which the direct access is not easy and until
now, the intrauterine devices are only used in the art of
contraceptive methods. However, the uterine cavity may be used for
several other purposes with an appropriate intrauterine device,
notably for treating directly the uterine wall (endometrium and/or
myometrium), for temporary in vivo embryo/egg incubation and as an
appropriate administration way to reach the general blood system.
The present invention provides with means in this connection.
[0002] The present invention relates to a retrievable intrauterine
(uterine cavity or Fallopian tube) device notably for implantation
of gametes or embryos into the uterus (or tube) allowing to perform
in vivo and in utero (or intraFallopian tube) fertilization and/or
preimplantatory development in assisted reproductive technology
(ART).
[0003] The invention also relates to a similar intrauterine device
for implantation of genetically modified cell lines by genes
transfection into the uterus in order to deliver molecules near the
endometrium without systemic effect and permitting to modify and
prepare more specifically the endometrium before embryo transfer
after ART or natural conception, or in contrary to avoid any
pregnancy (contraception) as standard IntraUterine Device
(IUD).
[0004] The invention also relates to a similar intrauterine device
allowing the delivery of various active elements within the uterine
cavity.
[0005] In particular, said active elements can be both
gametes/embryos and genetically modified cell lines by genes
transfection for bioactive factors secretion in order to improve
the uterine environment for in vivo incubated gametes/embryos
(embryos possibly resulting from cloning or any other
technique).
[0006] The present invention can be applied to any mammalian
species.
[0007] According to a first aspect, the invention concerns
reproductive medicine in particular the in vitro fertilization
(IVF) in assisted reproductive technology and drug delivery from
the uterus using an intrauterine device associated to cell
encapsulation technology.
[0008] Since the introduction in 1978, IVF has become the preferred
procedure for addressing most causes of infertility in humans. As a
part of the IVF process, the reproductive cells (oocytes and
spermatozoa) and the resulting fertilized oocytes (zygotes,
embryos) are treated according to specified procedures using in
vitro culture media adapted to each specific step in the
procedure.
[0009] A standard in-vitro fertilization (IVF) comprises the
following steps: [0010] Maturation (Oocyte recruitment) To ensure
maturation of more than one oocyte, the women are treated with
hormones prior to the actual fertilization procedure. Usually the
woman is treated for 14-21 days with a element (GnRH agonist) which
will disrupt then normal hormonal control signals between the brain
(hypothalamus and pituitary) and the ovary. Thereafter relatively
large doses of FSH (Follicle Stimulating Hormone) are administered
for 10-20 days depending on ovarian response. FSH will stimulate to
maturation of many follicles each containing an oocyte. When the
oocytes are ready for ovulation, human Chorionic Gonadotrophin
(hCG) is administered to finalize oocyte maturation. [0011]
Aspiration of oocytes After maturation in vivo, the oocytes are
collected from the woman's ovaries using ultrasound guided
follicular puncture. [0012] Fertilization and embryo culture In
vitro fertilization is obtained by adding spermatozoa to the
oocytes (In vitro fertilization IVF) or by microinjecting one
spermatozoon into each mature oocyte (Intracytoplasmatic sperm
injection-ICSI). The fertilized oocytes are cultivated in IVF media
outside the genital tract for two to five days. [0013] Embryo
transfer Alter two to five days of embryo in vitro culture a few
embryos are selected and transferred into the woman's uterus using
a thin catheter. The ultimate goal of in vitro fertilization and
embryo culture is to provide high quality embryos that are capable
of continuing normal development and results in live births. [0014]
In vitro culture for preimplantatory embryo development Despite
nearly 20 years of treating patients with IVF and more recently
intracytoplasmic sperm injection (ICSI), implantation rates per
embryo transferred remain low, on average about 20%.
[0015] Most IVF centers around the world perform the embryo
transfer at day 2 or 3 which means 3 or 2 days before the
physiological implantation time. Recent development in the field of
embryo physiology and metabolism have led to the formation of new
sequential serum-free culture media designed to simulate the
dynamic environment as the embryo travel along the reproductive
tract (Gardner et al., 1996). Sequential culture media systems
(G1.2/G2.2) have the highest rate of blastocyst formation which
remains low at 50% (Gardner et al., 1998).
[0016] The rationale for culturing embryos to the blastocyst stage
is that it allows selection for transfer of embryos with proven
developmental capacity. Furthermore, the transfer of a blastocyst
into the uterus is physiologically closer to the in vivo situation
than transferring an early cleaving embryo that would normally be
present in the Fallopian tube and has less risk to be expulsed from
the uterus because of a reduced time before embryo
implantation.
[0017] Most recent papers have shown the beneficial effect of in
vitro coculture of the zygote with human endometrial epithelial
(Simon et al., 1999) and/or stromal (Barmat et al., 1998) cells on
the number of blastomeres per preembryo, the rate of development to
the morula-blastocyst stage, the rate of spontaneous hatching and
the percentage of cytoplasmic fragments, and implantation rate as
recently described by Spandorfer et al., 2002.
[0018] In the report of the year 1994 of assisted reproductive
technology activities in the United States and Canada, gamete
intrafallopian transfer (GIFT) and zygote intrafallopian transfer
(ZIFT) have the higher rate of clinical pregnancy compared to in
vitro fertilization. More recently, Levran et al., in 2002
demonstrate that zygote intrafallopian tube transfer improve the
outcome in repeated implantation failure compared to a transfer of
blastocysts after standard IVF.
[0019] These findings would suggest that the presence of zygote
into the upper genital tract may be important for the embryo
development profiting of all known and unknown growth factors
present in the uterine fluid and its potentiality to invade the
endometrium during the implantation process.
[0020] Furthermore, in bovine studies have recently demonstrated
that in vivo produced embryos were less alterated compared to in
vitro ones, with more intercellular communication devices (Boni et
al., 1999) and more mature mitochondria in particular (Crosier et
al., 2000).
[0021] All the above mentioned studies confirm that in vitro embryo
preimplantatory development is far to be optimal instead of all
efforts to optimize culture media by mimic uterine fluid
characteristics.
[0022] The present invention provides with a novel method in ART
using the cell encapsulation technology in order to notably permit
to gametes and/or preimplantatory zygotes/embryos in IVF programmes
(or cloning for all other mammalian species) to benefit from a
temporary natural incubation into the uterus.
[0023] Furthermore, a controlled in time of the intrauterine
incubation may lead to a better quality of embryo development and
consequently to a higher implantation rate, and it may have an
economical advantage with a lower cost/benefit to the standard IVF
procedure above all if blastocyst transfer is generalized in ART
units.
[0024] According to a second aspect, the invention relates to the
in utero delivery of active elements by the implantation of
tissues, cells or cell lines, possibly genetically modified,
notably for cellular therapy.
[0025] In all mammalian species the success of implantation is
related to a perfect crosstalk between a good quality embryo and a
receptive endometrium.
[0026] In the domain of assisted reproductive technology (ART),
clinicians are limited in the control of the complex events of
endometrium receptivity by endocrinological treatment by the
administration of 17beta estradiol and progesterone to mimic the
physiological sequential follicular and luteal phases.
[0027] In the basic science literature, most studies are now
focusing on the paracrinology of periimplantation between the
embryo and the endometrium using in vitro and in vivo models as
knock-out mouses lacking the interest molecule. For instance, it
has been shown that in integrin beta1-deficient mice (Fassler and
Meyer, 1995) and female mice with a null mutation of interleukin-11
receptor alpha chain (Robb et al., 1998) embryos failed to
implant.
[0028] However, in rodents it is established that direct contact
between the embryo and the endometrium is not necessary (Shiotani
et al., 1993). Whereas embryo implantation rate is remained stable
and low since two decade we can say that endocrine manipulation
with vaginal progesterone and hCG systemic administration and
finally 17beta oestradiol are far to be optimal and quite far to
the complex paracrinology of embryo implantation processes.
[0029] The present invention provides a novel concept in ART using
the cell encapsulation technology in order to implant cell lines
genetically modified secreting a molecule of interest near the
endometrium temporary before embryo transfer in IVF programmes.
[0030] Such an intrauterine molecule(s) or "drug" delivery, closer
to the target tissue permits a better and more selectively
preparation of the endometrium and a use of small molecules with
short half time or molecules with secondary effects forbidden in
systemic administration.
[0031] According to a third aspect, the invention relates to an
implantable/insertable device to deliver agents capable of
preparing the uterus before receiving embryo(s) before implantation
into the wall of the uterus or for treating the uterus. With
respect to this embodiment, certain or most drugs, nutrients,
vitamins, amino acids, fatty acids, peptides, proteins and the like
capable of stimulating the uterus specifically with respect to
preparation of embryo implantation, or any other therapeutical
agent can also be easily delivered by the device of the invention
containing polymers or cells releasing these agents. In fact, the
retrievable intrauterine device of the invention for drug delivery
has specific advantages for women who can not receive drugs
normally given by injection or other delivery routes. In addition,
delivery of agents through the uterine wall may be an optimal means
for treating forms of reproductive system cancers and/or other
reproductive diseases, as well as any uterine diseases.
[0032] The invention notably provides a cell encapsulation
intrauterine device for in vivo and in utero gametes fertilization
and/or embryo preimplantatory development with a control of time
(from several minutes to 48, 72 h) where the uterus play a role of
"natural incubator" before the definitive embryo transfer in IVF
programmes.
[0033] Thus, the cell encapsulation intrauterine device of the
invention is a novel and modified intrauterine device, similar to
contraceptive IUD as described in U.S. Pat. No. 3,628,530 by Jerome
Schwarz in 1969 and in U.S. Pat. No. 3,516,403 by Rene Cournut in
1967, which has, according to the present invention, to permit a
temporary introduction of gametes or embryos associated or not to
other somatic cells (in vivo coculture) into the uterine cavity and
its retrieval after a defined time by using a cell encapsulation
technology as described in WO 91/00119 by Thomas Mandel et al. in
1989, WO 01/64185 A2 by Newman and Kram in 2000 and in U.S. Pat.
No. 5,158,881 by Aebischer et al. in 1990 and U.S. Pat. No.
6,054,142 by Li et al. in 1996.
[0034] In fact, the present invention concerns a retrievable
intrauterine device for placing within the uterine cavity one or
more encapsulated element(s) capable of having interactions with
the uterine fluid comprising an intrauterine device loaded with
encapsulated elements.
[0035] According to the present invention, the term "element(s)"
means any organic or inorganic, cellular or molecular, natural or
synthetic compound(s) or substance(s).
[0036] The term "loaded" encompasses not only the case where the
intrauterine device of the invention is the support of said
encapsulated element which can be for example be adsorbed on the
surface of said device, but also the case where said elements are
contained within the device of the invention. According to a
preferred embodiment, the retrievable intrauterine device of the
invention is provided with at least one housing in which said
encapsulated element(s) is/are loaded. Different elements may be
mixed in the same housing or placed in separate housings.
[0037] The expression "capable of having interactions with the
uterine fluid" encompasses not only the case where said elements
are delivered by the device of the invention in the uterine fluid
in order to produce an effect on the wall (endometrium and/or
myometrium) of the uterine cavity (first case), but also the case
where said elements are staying within the device and have known or
unknown exchanges or interactions with the uterine fluid and/or
with the uterine wall (endometrium and/or myometrium) (second
case).
[0038] According to a first aspect of the "first case", said
elements may be not only agent(s) capable of treating the uterus
but also agent(s) capable of treating any pathology. In fact,
agent(s) capable of treating the uterus will be delivered from the
device of the invention into the uterine fluid and will have an
effect directly on the wall of the uterine cavity. This kind of
agent is selected from the group comprising agent(s) capable of
preparing the uterine wall for optimal embryo implantation,
agent(s) capable of preparing the uterine wall for optimal egg
culture, agent(s) capable of therapeutically treating the uterus
and contraceptive agent(s).
[0039] The advantage of such a system allows using agents capable
of having a direct effect of the target organ in avoiding the
systemic way.
[0040] Furthermore, according to a second aspect of the "first
case", the device of the invention can allow the delivery of agents
capable of treating any pathology by the in utero administration.
In other words, this kind of agent will be delivered in the uterine
fluid, will then enter into contact with the wall of the uterine
cavity and will pass through the venous system of the endometrium
before passing through the general blood system. In other words,
the in utero administration is a possible way of administration of
any drug likely to treat any pathology in connection with any
organ.
[0041] Among the elements likely to be loaded in the retrievable
intrauterine device of the invention, one must also cite tissues,
cells or cell lines secreting one or more agent(s) (for cellular
therapy), somatic cells, stem cells (totipotent cells), recombinant
viruses as gene transfer vehicle (for gene therapy), sens or
antisens mRNA sequences, male and/or female gametes, fertilized
oocyte (two pronuclei cell), unfertilized egg and any combination
of the above elements.
[0042] The above "second case" concerns fertilized oocyte (two
pronuclei cell) and unfertilized egg where the uterus plays the
role of a natural incubator allowing the culture of said embryo or
egg in a natural medium in lieu of an in vitro medium. In this
case, no element loaded in the intrauterine device of the invention
is delivered into the uterine fluid and the expression "having
interactions with the uterine fluid" encompasses the situation
where the embryo or the egg has exchanges or interactions with the
environmental medium constituted by the uterine fluid and the
uterine wall.
[0043] According to a preferred embodiment of the present
invention, the at least one agent secreted by tissues, cells or
cell lines is selected from the group comprising drugs, hormones,
nutrients, peptides, proteins, antibodies, trophic factors, growth
factors, lymphokines, cytokines, enzymes, blood coagulation
factors, angiogenesis factors, analgesics, neuro-transmitters,
neuromodulators. In this connection, the present invention
encompasses the case where tissues, cells or cell lines secreting
agents likely to be loaded in the device of the invention can be
genetically modified in order to obtain the secretion of the
desired product.
[0044] According to another preferred embodiment of the invention,
the agent capable of therapeutically treated the uterus is selected
from the group comprising anti-inflammatory agents, amino acids,
fatty acids, antibodies, trophic factors, growth factors,
lymphokines, cytokines, enzymes, proteins, peptides, blood
coagulation factors, angiogenesis factors, analgesics,
neurotransmitters, neuromodulators, anxiolytics, antidepressants,
antibiotics, sens or anti-sens mRNA sequences, recombinant viruses
as gene transfer vehicle.
[0045] According to another embodiment of the present invention,
the element loaded in the retrievable intrauterine device of the
invention is capable of treating cancers, forms of reproductive
system cancers, reproductive diseases and uterine diseases such as
endometriosis, adenomyosis, bleeding disorders and various
infections (non limitative list).
[0046] In fact, the device of the invention presents a number of
advantages: said device is not likely to cause any trouble to the
uterus due to the fact that it is not implanted within the uterine
wall, it does not require-surgery or anesthesia to be inserted and
it can be inserted in a completely ambulatory fashion. Furthermore,
elements may be delivered in phase with the natural menstrual cycle
and in association with hormone dependent gene expression systems
controlling drug delivery from the device. It is also very
important to note that, as the percutaneous way of administration,
the in utero administration allows to avoid the first hepatic
passage of the administered element(s). This causes less toxicity
and said element(s) present(s) a better bio availability. Finally,
said device can be rapidly retrieved with surgical
intervention.
[0047] FIG. 1 illustrates an example of device according to the
present invention (with 1: capsule, 2: support zone A--distal
part--and support zone B--proximal part, 3: plunger, 4: protective
tube, 5: membrane or valve for disclosure, 6: wings, 7: removal, 8:
silicone tether).
[0048] As example, the device of the invention comprises:
[0049] 1. a capsule [0050] non-biodegradable [0051] semi permeable
[0052] polymeric material (i.e. polyethersulfone or PES.
Source=Akzo Nobel Faser AG, Wuppertal, Germany) hollow fibers such
as polyacrylates (including copolymers), polyvinylidienes,
polyurethanes [0053] pores: has to be adapted for an optimal
environment quality with a permeability from small to large
molecules present into the uterine fluid [0054] Pore-size: from
0.005 .mu.m=molecular weight cutoff of 150 kDa to 280 kDa [0055]
Pore-structure: microporous or any adapted structure [0056] Outer
diameter=adapted to the optimal size to go through the cervix into
the uterine catheter (i.e. 700 .mu.m) [0057] inner diameter=adapted
to the size of egg or embryo (i.e. 500 .mu.m), around 5 times the
egg or embryo size (egg size with the corona radiata=200 .mu.m in
human), or adapted to the size of the molecules to be delivered,
[0058] length adapted to the uterine cavity avoiding endometrial
lesions and any deleterious factor by uterine distension and
finally its expulsion (i.e. about 1.5 cm) [0059] cylindric shape or
any adapted shapes for the uterine cavity taking into account the
above-mentioned point.
[0060] 2. two supports, zones A (distal) and B (proximal) as
described for IUD in U.S. Pat. No. 3,628,530 by Schwarz with the
following modification: [0061] distal part (A) closed by
acrylate-based glue, Luxtrack LCM 23 (Ablestik, USA) or any other
system as cap, proximal part (B), hollow and openable with a one
sens direction (on the inside) valve. [0062] zone A support
comprises two wings or any other devices (it can comprises
asymmetrical wings in a similar material than the support covered
by a thin hydrophilic material which becomes thicker at fluid
contact) in a similar material or others than the support
permitting a stable position of the device into the uterus. [0063]
zone B support comprises a silicone tether to attach the removing
thread in order to retrieve the device
[0064] 3. a plunger
[0065] 4. an operating handle being coupled with the plunger
[0066] 5. a removing fiber for removing the intrauterine
encapsulating device
[0067] 6. an attachment device for attaching the removing fiber
ensuring immovable position of the intrauterine encapsulating
device with respect to the plunger
[0068] 7. a protective tube encompassing the plunger as described
by Lehtinen Matti et al. in 1994 (patent number CZ 286 820), but
with a plunger permitting the loading of gametes; embryos, drugs or
any other element with a standard pipette by the IVF biologist.
[0069] The present cell encapsulation intrauterine device may take
any shape which will accommodate gametes and/or embryos or any
other above mentioned element to be encapsulated using an adapted
micropipette for loading. A preferable implantable embryo culture
device is a tubular, selectively permeable membrane with an adapted
pore size in order to permit adequate nutrients transfer to the
embryo such as O2, proteins, growth factors and other known and
unknown factors released from the endometrium, having one end
through which gametes or embryos are loaded into cell compartment.
The controlateral end may then be permanently occluded with caps or
alternatively with an epoxy glue or sutures of a biocompatible and
non resorbable material like polypropylene.
[0070] Concerning the structure of the retrievable intrauterine
device according to the present invention, a retrievable hollow
fiber device with internal diameter, dimensions ranging from
100-10,000 microns and having a suitable means for attachment of
the device to uterine cavity (wall) is ideal as an egg/embryo
chamber for inter uterine incubation. The current device design
employed in connection with the present invention is a
Polyethersulfone membrane with MW cutoff of approximately 240,000
daltons, wall diameter of 100 microns and inner diameter of 472
microns. Wall thickness can range from 50-500 microns depending on
the composition, porosity, hydraulic permeability, pore size and
strength of the encapsulating material. Molecular weight cutoffs
can vary from 50,000 to >1 million molecular weight.
Furthermore, the encapsulating material can be composed of any
biocompatible material including Polyethersulfone, Pan-PVC or
expanded PTFE and formulated with laminated or single membrane
structures. The encapsulating device may or may not contain a
matrix or internal lining material. The length can range from 0.5
cm-5 cm or what ever will fit comfortably within the uterus. As
indicated above, one of the most important considerations for this
device are that it be capable of holding an embryo without damage
for incubation directly in the uterus to be fully retrievable. In
addition, the device has been designed with little or no tissue
reaction, with smooth surfaces such that it will be
implantable/insertable and retrievable without inducing
inflammatory or fibrotic reactions and or inappropriate uterine
wall tissue damage or scarring. Finally, the device has been
designed such that it has a means of remaining within the uterus
(i.e. small suture thread glued into the tip of the device for
attachment inside or outside the uterus) and that can be easily
retrieved (attached suture thread) at any time after uterine
implantation. Devices similar in composition to those used
currently by Modex Therapeutics (PES 5, PES 1, PES 10/10) hollow
fiber device and used in the past by Cytotherapeutics (Pan-PVC) are
ideal for such applications although any device, which meets this
general description will be appropriate for the contemplated
uses.
[0071] The device of the invention can be surgically implanted
passing through the cervix as a standard IUD for contraception into
the uterus and removed after a defined time of incubation.
[0072] However, the present invention may use as implantation site
the Fallopian tube permitted by a modification of the present cell
encapsulation intrauterine device for instance without the distal
part with the two wings. This implantation is more difficult in
access and need to use a surgical procedure as coelioscopy with
general anesthesia or culdoscopy with local anaesthesia. Such in
vivo intra Fallopian embryo culture is similar to GIFT or ZIFT
except the fact that using the cell encapsulation device of the
invention the incubation time is under control and an unlimited
number of embryos can be loaded which were retrieved and selected
for transfer after a simple flushing procedure.
[0073] Thus, zygotes or embryos at different stages (day 2 to 5)
can be loaded with a standard micropipette into the capsule of the
cell encapsulation intrauterine device of the invention after
standard IVF with or without ICSI.
[0074] The invention also relates to a method of preparing a
retrievable intrauterine device for placing one or more
encapsulated elements capable of having interactions with the
uterine fluid comprising the steps of: [0075] providing said
element under the appropriate form to be encapsulated, [0076]
providing a retrievable intrauterine device suitable for receiving
encapsulated element(s), [0077] loading said device with said
element(s).
[0078] The invention also relates to a method for placing one or
more encapsulated elements capable of having interaction with the
uterine fluid comprising the steps of: [0079] providing a
retrievable intrauterine device, [0080] implanting said device
within the uterus cavity for a determined duration.
[0081] According to a preferred embodiment of the invention, the
above method is carried out in the uterine cavity of a mammal
preferably selected from the group comprising bovine, ovine,
porcine and humans.
[0082] More particularly, the present invention may be carried out
according to the following process:
[0083] A. In Vivo Fertilization
[0084] Injection of prepared sperm and retrieval oocytes in the
encapsulating device for implanting into the uterus. After a
defined and controlled incubation time (2 hours par example) of in
vivo and in utero culture, encapsulated sperm and oocytes are
retrieved and zygotes and/or non fertilized oocytes are collected
after a simple flushing procedure. Selection of zygotes for
cryopreservation or in vitro culture of the remaining embryos to be
transferred at day 3.
[0085] B. In Vivo Preimplantatory Embryo Development
[0086] Conventional in vitro fertilization of prepared sperm and
retrieved oocytes. Injection of several embryos at different stage
of development (i.e 6-8 cells) in the cell encapsulation device
permitting the implantation into the uterine cavity during a
controlled time (i.e 48 hours). After removal from the uterus the
encapsulating system device embryos at the blastocyst stage are
flushed from the device and transferred into the uterine cavity
using a conventional transfer catheter or delayed in a further
cycle after freezing.
[0087] C. In Vivo Embryo Assisted Hatching
[0088] Recently, it has been demonstrated that the process of
blastocyst hatching as well studied and described in in vitro
programmes has been erroneously accepted in rodents as representing
a natural event (Gonzales et al., 2001). Indeed, it seems in
hamster species that the uterine contribution in vivo to blastocyst
escape from the zona pellucida, consisting of proteinases secreted
from the uterus, is the primary mechanism for zona loss in utero,
whereas the in vitro lytic activity is secondary to the invasive
behavior of trophectoderm.
[0089] Taking into account such surprising results, the present
invention using the intrauterine cell encapsulation device of the
invention to incubate temporary embryos can be used to perform a
novel method for assisted hatching: the in vivo embryo assisted
hatching.
[0090] On this basis, in vivo and in utero culture of the gametes
and/or preimplantatory embryos with a time control using the cell
encapsulation device of the invention will permit a real dialogue
at the embryo-maternal interface with a paracrine action of several
known but also unknown factors from endometrium (or from the tube
in case of intraFallopian implantation) and embryos important for
the optimal embryo development which will lead to a better success
of implantation process in ART programmes.
[0091] Other scientists have described gametes or embryos
encapsulation (Loi et al., 1992; Nebel et al., 1993). However, they
all used biodegradable materials (sodium alginate) and have as
objectives to eliminate several problems associated with the
procedure of embryo transfer (trauma) and to improve embryo
protection before implantation and to protect the free zona
pellucida of the embryo (Cosby at al., 1990; Adaniya et al.,
1993).
[0092] To the best of the knowledge of the inventor, the novel
concept described in the present invention permitting a natural
incubation of gametes and/or embryo by using the cell encapsulation
device of the invention has never been published or proposed by
scientists.
[0093] The invention also provides with a method using a cell
encapsulation device with genetically cell lines by genes
transfection as described in U.S. Pat. No. 4,686,098 by Kopchick et
al. in 1984 and U.S. Pat. No. 4,892,538 by Aebischer et al. in 1957
modified in order to be implanted into the uterine cavity which has
never been published by scientists before the present
invention.
[0094] This novel device as mentioned above permits a novel method
to deliver from the uterine cavity molecules near the endometrium
without systemic effect and permit to modify and prepare more
specifically the endometrium before embryo transfer after in vitro
fertilization in ART or natural conception, or in contrary to avoid
any pregnancy (anti-implantatory, anti-fertilization) as standard
IUD.
[0095] This novel concept of bioactive factors delivery into the
uterine cavity may lead to a better understanding of the specific
paracrine effect on the endometrium tissue and may permit in the
near future the development of a novel and complementary cellular
therapy approach to modulate and prepare the endometrium for embryo
implantation in ART.
EXAMPLE 1
In Vivo and in Utero Embryos Culture in a Mouse Model
[0096] The purpose of this experiment was to evaluate the ability
to perform in vivo and in utero preimplantatory embryo development
using a modified semi-permeable hollow fiber as a capsule in a
mouse model.
[0097] Zygotes were obtained using a standard ovarian stimulation
protocol in 4- to 5-week old prepubertal females using 5 IU PMSG,
which corresponds to day 1 of the procedure (Folligon, Veterinaria)
and 5 IU human chorionic gonadotropin, i.p. (Choluron,
Veterinaria), day 3, at 17:00, 48 h apart, in order to induce
superovulation.
[0098] Females were caged with CBAxC57B1 males at the time of the
HCG injection (day 3).
[0099] Embryos at 6-8 cells were collected at day 5 after caging
males and females and cultured either in vivo (group 1) or in vitro
using a sequential medium culture (group 2). Only two female with a
copulation plug were operated and used for this experiment.
[0100] Females were killed by cervical dislocation and laparotomy
was performed to exteriorize the uterine horn and the tube in order
to collect embryos.
[0101] Transfer of 6-8 cells embryos into the right or the left
horn were performed at day 3 of two pseudopregnant females
(recipients).
[0102] Group 1 embryos (in vivo culture) were loaded into a
modified hollow fiber device according to the invention (semi
permeable polyethersulfone (PES) hollow fibers, Source=Akzo Nobel
Faser AG, Wuppertal, Germany) with an outer diameter=680 um and
inner diameter=480 um of 0.5 cm length, attached to a 6.0
sterilized and non resorbable surgical thread by using a fine glass
pipette under microscopic visualization.
[0103] A dorsal laparotomy was performed under ether anaesthesia
according a standard procedure in order to exteriorize the right or
the left horn and implant the intrauterine device with encapsulated
zygotes into the lumen of right or left horn.
[0104] After fixing surgically the intrauterine device with
encapsulated zygotes the horn was replaced in its anatomical
position and hooks were used to close the skin.
[0105] After a period of 48 h transferred females were killed by
cervical dislocation and laparotomy was performed to exteriorize
the left or right horn containing the intrauterine device and to
retrieve it.
[0106] Embryos were collected after cutting both distal part of the
device and a capsule cavity flushing with culture medium.
[0107] Table 1 shows the results of two experiments comparing the
in vivo and in vitro (as control) development of 6-8 cells embryos
after 48 h culture.
[0108] In group 1, all encapsulated 6-8 cells embryos continued
their development into the uterus similar to the control group.
However, a delay was noted in development compared to the in vitro
embryos culture.
[0109] This example shows for the first time that intrauterine
device with encapsulated embryos may allow their natural incubation
into the uterus with a possible development at least similar to the
conventional in vitro culture.
[0110] From the available literature, IUD has ever been considered
as hostile to fertility and always related to contraception. The
presence of a foreign body in the uterine cavity is known to
interfere with reproduction in all species. However, the affected
steps of reproductive processes are far to be clear in literature.
It appears that it may vary according to species. It is generally
accepted that IUD induce a local inflammatory reaction in the
endometrium. Whereas in mouse and rat this chronic infiltration of
polymorphonuclear seems to be related to bacterial infection and to
transforms the uterine endometrium in an hostile environment with
embryo toxic secreted factors (Parr et al., 1967), 20 years after,
Alvarez et al. (1988) showed in women that IUD may affect the
fertilization before that the blastocyst enters the uterine
cavity.
[0111] The above results demonstrating a mouse embryo development
without degeneration are against the generally accepted scientific
opinion that in animal models the uterus itself kills embryos by
releasing several toxic factors.
[0112] The delay in development as shown in in vivo encapsulated
embryos may be explained to the unoptimal size of pores or inner
lumen diameter leading to a lower concentration of nutrients around
it. Furthermore, the time itself which was sometimes several
minutes in room temperature to implant the IUD with encapsulated
embryos into the uterus before in vivo culture may explain thermic
and gazous shockes with deleterious effect on preimplanted
embryo.
EXAMPLE 2
Intrauterine Erythropoietin Delivery
[0113] Intrauterine erythropoietin (Epo) delivery using a modified
and invented encapsulated mouse Epo secreting mouse C2C12 cells
decrease apopotosis in endometrium and increase blood hematocrit
suggesting a direct effect of EPO on nearby endometrium tissue and
systemic effect of Epo delivered from the uterus.
[0114] Erythropoietin is produced by the kidney in adults and the
liver in fetuses. It is a key factor for regulating erythropoiesis
by stimulating proliferation and differentiation of late erythroid
precursor cells. It has been shown recently that the brain has a
paracrine Epo/EpoR and that Epo may prevent neuronal apopotosis
after cerebral ischemia (Siren et al., 2001). Interestingly, Epo
seems to be implicated in uterine angiogenesis.
[0115] Taking into account the above mentioned physiological effect
of Epo, the evaluation of the effect of Epo on endometrium by
delivering Epo from the uterus using the cell encapsulation device
of the invention and the evaluation of the cell viability in this
new implantation site for a device the uterine cavity by measuring
the blood hematocrit have been made.
[0116] Mouse C2C12 myoblast cells were transfected with a plasmid
containing the mouse Epo cDNA and a mutated dihydrofolate reductase
(DHFR) gene for gene amplification upon administration of
increasing doses of methotrexate.
[0117] Epo secreting cell lines were loaded into polyethersulfone
microporus hollow fibers in order to be implanted into the uterine
cavity. Characteristics of this novel invented cell encapsulation
device are similar to the description in example 1.
[0118] A total of 14 devices were used implanted in a blinded
fashion in the present experiment, group 1: mEpo-C2C12 (n=7) with
Epo secretion and group 2: mEpo-C2C12 control cells (n=7) without
Epo secretion.
[0119] At day 14, according to a standard accepted protocol for
animal sacrifice, the uterus was removed and capsule retrieved in
order to be pulsed for mEPO output. The uterus was fixed in 10%
formol for immunohistochemistry and TUNEL assay to test
apoptosis.
[0120] A significant increase blood hematocrite in group 1 with
intrauterine Epo delivery (59.4+/-6.8) compared to group 2 without
Epo delivery (45.7+/-2.9), p.ltoreq.0.005 is noted after 14 days of
intrauterine Epo delivery in 14 female mice (data not shown
here).
[0121] It is noted that Epo, similar to its effect recently
described in neuronal cells, decrease apoptosis in endometrium
tissue.
[0122] The endometrium is thus a new target of Epo with a
modulation of apoptosis.
[0123] Transfected cell line encapsulated in microporous hollow
fiber are viable after 14 days of intrauterine incubation which has
never been tested as implantation site for cell encapsulation
device before.
[0124] Epo secreted by transfected cell line go across the
microporous wall of the device and is resorbed through the
endometrium and through the systemic circulation according to the
significant increase of blood hematocrit intreated animals.
[0125] These results confirm that i) the uterus may be an excellent
natural incubator confirming the results of example No 1 for embryo
development and ii) drug delivery may use in woman or female
animals the uterine cavity as implantation site of a cell
encapsulation system.
TABLE-US-00001 TABLE 1 t 48 h in vivo in vitro 6-8 6-8 Experiments
cells morula blastocysts cells morula blastocysts No1 n = 20 n = 10
6-8 cells N = 30 0 16 2* 0 0 9.sup..dagger-dbl. No2 n = 12 n = 10
6-8 cells N = 22 0 6 1 0 2 8.sup. *early blastocysts, no retrieval
after capsule flushing of 2 embryos .sup..dagger-dbl.1 early
blastocyst, 1 embryo losted
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