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`PCT/US2011/026741
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`d.
`
`At 248d, immersion of the tissue in 0.1 - 25% glutaraldehyde (filtered to limit
`
`oligomeric content) for between about 3 days to 5 weeks, and more preferably, for
`
`between about 3 days to 4 weeks, and more preferably yet, for between about 3 weeks to
`
`4 weeks, at Oto 37°C, and more preferably, 0.25% glutaraldehyde for 7 days at 4°C; or
`
`5
`
`e.
`
`At 248e, immersion in the tissue in one of the above formalin, glutaraldehyde, or
`
`oligomeric filtered glutaraldehyde solutions together with added amino acids, lysine
`
`and/or histidine, wherein the concentration of the amino acids, L-lysine or histidine, used
`
`as an additive to the fixative is in the range of about 100 - 1000 milliMolar, with a
`
`preferred value of about 684 mM.
`
`10
`
`In addition to the foregoing, combinations of the processes listed above may be
`
`performed, including: step a followed by step b; step a followed by step c; and step a followed
`
`by step d.
`
`As those skilled in the art will appreciate, heat-shrink testing may be conducted on tissue
`
`samples to correlate the effectiveness of protein cross-linking. Here, results of heat-shrink
`
`15
`
`testing performed on one or more samples of tissue prepared in accordance with at least one
`
`embodiment using formalin showed that the tissue had a shrink temperature of 90°C. This
`
`compares favorably with samples prepared using glutaraldehyde, wherein the shrink temperature
`
`was 80°C. Accordingly, formalin is a suitable variant of fixation. It is noted that formalin was
`
`generally abandoned by the field, largely because of material properties that were unfavorable
`
`20
`
`and because of inadequate or unstable protein cross-linking. Such problems have been
`
`overcome through the pretreatments described herein, allowing production of tissue with
`
`strength, pliability, and durability in a relatively thin membrane. When used in a prosthetic
`
`implant, such as a heart valve, the tissue characteristics imparted by the tissue preparation
`
`process facilitate formation of a construct having a relatively low-profile, which also thereby
`
`25
`
`facilitates dry packaging of the prosthetic implant. The same advantages are also achieved using
`
`the pretreatments when using a glutaraldehyde process.
`
`Referring still to Fig. 2B, after fixation for collagen cross-linking at 248, an alcohol post(cid:173)
`
`fixation treatment at 252 is preferably performed by rinsing the tissue in distilled water at 256,
`
`and then at 260 rinsing the tissue in 25% isopropyl alcohol for between about 30 minutes to 14
`
`30
`
`days or more at between about Oto 37°C, and more preferably, for at least about 7 days at 20°C.
`
`At 264, the tissue undergoes a rinsing with distilled water.
`
`In accordance with at least one embodiment, treatment of the tissue, including from the
`
`time of harvest to the time of implantation or grafting, does not include contact and/or exposure
`
`to a polymer to infiltrate and/or encapsulate tissue fibers of the tissue.
`
`10
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1551 of 2218
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`
`Referring now to Fig. 3, the drying process at 300 is performed after the tissue
`
`preparation at 200. Thus, in accordance with at least one embodiment, the tissue is dried under a
`
`load. More particularly, for the tissue drying at 304, the tissue is placed minimally stretched flat
`
`(that is, stretchedjust enough to eliminate visible wrinkles and bubbles) on a flat surface (e.g., a
`
`5
`
`polymer or acrylic sheet) at 308, and held fixed at its edges at 312. Optionally, the joined tissue
`
`and underlying sheet are then set in a slight curve. The tension maintains the substantially flat
`
`structure of the tissue as it dries, thereby mitigating or preventing excessive shrinkage,
`
`wrinkling, and/or curling at the edges, and also making the rate of drying more uniform across
`
`the surface of the tissue because of the surface tension between the plate and the tissue.
`
`10 Alternatively, the tissue is dried while compressed between acrylic plates. When drying the
`
`tissue, the temperature is held at between about 4 to 37°C, and more preferably, between about
`
`20 to 37°C (i.e., approximately room temperature to normal human body temperature), and more
`
`preferably, at about 20°C. At 314, the drying process is performed in substantially dark
`
`conditions (i.e., substantially no visible light) for between about 6 hours to 5 days, and more
`
`15
`
`preferably, for about 72 hours. By way of example, the tissue is dried in dark conditions at a
`
`temperature of about 20°C for between about 6 hours to 5 days, and more preferably, for about
`
`72 hours. As those skilled in the art will appreciate, drying the tissue while the tissue is
`
`compressed between plates requires a longer period of time.
`
`In at least one embodiment, after drying, the tissue lots are inspected at 316, such as by
`
`20
`
`stereomicroscopy, to identify and discard those with defects or discontinuities of the fiber
`
`matrix. If desired, the preferential fiber direction for each piece may be identified to determine a
`
`particular orientation, for example, to determine the free edge of the pieces that will form valve
`
`leaflets for a heart valve. Depending upon the size (i.e., the area) of the tissue being prepared
`
`and the size of tissue needed for a given implant, the tissue may be trimmed or otherwise sized
`
`25
`
`in optional sizing at 320, such as by cutting the tissue into an appropriately sized and shaped
`
`sheet for implant formation and/or manipulation. Preferably, cutting of the tissue membrane is
`
`oriented so that the resulting free edge is parallel to the preferential fiber direction of the tissue
`
`membrane. Optionally, the free edge may also be cut with a parabolic or other curved profile to
`
`compensate for any attachment angles in order to increase the total contact surface between the
`
`30
`
`tissue membrane and any associated frame or other structure. This approach minimizes
`
`weaknesses in the operating margins of the tissue assembly and advantageously distributes the
`
`principal loading forces of the operating implant along the long axis of the collagen fibers. As a
`
`result, the tissue is resistant to surface fracture and fraying.
`
`As shnwn in Fig. 3, optional sizing at 320 is performed after the drying at 304 and
`
`35
`
`inspection at 316. A rectangular shaped piece of tissue 400 is shown in Fig. 4. The tissue 400
`
`11
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1552 of 2218
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`

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`
`may be manipulated for use in a variety of prosthetic implants and grafts.
`
`As mentioned above, tissue prepared by the methods described herein has been
`
`successfully prepared by rinsing with 25% isopropyl alcohol for a period of 7 days, and after the
`
`further treatment steps described herein, provided an ultimate tensile strength of greater than 25
`
`5 MegaPascals. Here, the combination of tissue pliability and tensile strength is sought for
`
`purposes of producing a material having property characteristics suitable for being physically
`
`manipulated to form prosthetic implants, such as a tissue leaflet assembly for a heart valve or a
`
`ligament, while providing a tissue material that will operate properly once implanted. These
`
`techniques are intended to conserve and preserve collagen fibers, minimize damage to the tissue
`
`10
`
`and improve tissue characteristics. The preparation and fixation techniques produce tissue
`
`membrane material that may be rendered and used at lesser thicknesses than typically rendered
`
`in the prior art. Thinner membranes are more pliable, but with conventional tissue preparation
`
`techniques the tensile strength of the tissue is sacrificed. Advantageously, the preparation
`
`techniques described herein have produced membranes that have as much as three times the
`
`15
`
`tensile strength of a commercial product of the prior art. This achieved strength is thus desirable
`
`for providing a tissue assembly having a low profile with appropriate durability, even in a
`
`substantially dry state. More particularly, the tissue possesses a relatively high tensile strength.
`
`By way of example and not limitation, testing has shown that embodiments of tissue prepared as
`
`described herein provide a tissue having a tensile strength of approximately three times the
`
`20
`
`tensile strength of current pericardia! valve tissue, such as on the order of approximately 25
`
`MegaPascals, thereby providing about 2,000 times the physiologic load strength for valve tissue.
`
`Moreover, testing of an embodiment of an implantable prosthetic heart valve made with tissue
`
`prepared as described herein and under a static load of greater than approximately 250 mmHg
`
`showed less than approximately 14% leakage, wherein such results are generally considered
`
`25
`
`superior to surgical tissue valve prostheses.
`
`With reference to Fig. 5, stress-strain curve results for five different tissue samples
`
`prepared in accordance with an embodiment are shown. For the testing results shown, the yield
`
`stress or ultimate tensile strength was obtained by attaching strips of tissue fixed at the ends in a
`
`linear force tester and increasing the length by 0.3 mm/sec while recording resultant force
`
`30
`
`(tension) until the material ruptured or separated entirely; these measurements were then used to
`
`calculate the stress-strain curves depicted in Fig. 5. As illustrated in the graph, the yield stress
`
`or ultimate tensile strength of the various tissue samples varied from about 30 to about 50
`
`MegaPascals. More particularly, for each curve shown in Fig. 5, the testing procedures were the
`
`same. That is, each of the curves shown pertain to separate pieces of tissue that were subjected
`
`35
`
`to the same test. The results show a minimum ultimate tensile strength of 30 MegaPascals, with
`
`12
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1553 of 2218
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`

`WO 2011/109433
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`
`a range up to 50 MegaPascals. Accordingly, the illustrated test results demonstrate consistency
`
`of the ultimate tensile strength results for the tissue treatment process.
`
`It is to be understood that the tissue generated from one or more of the tissue preparation
`
`procedures described herein may be used for a variety of devices or uses, and that use in a
`
`5
`
`prosthetic heart valve is but one possible application for utilizing the tissue. For example, the
`
`tissue may be used in a shunt, or as graft material for repair or modification of one or more
`
`human organs, including the heart and its blood vessels. By way of further example, the tissue
`
`may be used as a pericardial membrane patch for repair of congenital heart defects. The tissue
`
`also has application as a prosthetic tissue in tendon and ligament replacement, and as a tissue
`
`10
`
`product for wound management. Moreover, for use in a prosthetic heart valve, the tissue may be
`
`configured in a variety of ways and attached to a frame in a variety of ways. In addition, a
`
`plurality of separate tissue pieces may each be connected together, such as by suturing, to form a
`
`larger composite of treated tissue material. Thereafter, whether the prosthetic implant or graft is
`
`made of a folded tissue assembly or a plurality of separate tissue pieces, the resulting prosthetic
`
`15
`
`implant or graft may then be further manipulated for treatment of a patient.
`
`In at least one embodiment, tissue generated from one or more of the tissue preparation
`
`procedures described herein may be used to form a prosthetic implant that includes a stent,
`
`frame, bone screw or other fastening or anchoring mechanism. In yet other embodiments, tissue
`
`generated from one or more of the tissue preparation procedures described herein may be used to
`
`20
`
`form a prosthetic implant or graph that does not include a stent, frame, bone screw or other
`
`fastening or anchoring mechanism. Tissue generated from one or more of the tissue preparation
`
`procedures described herein may be may be packaged for delivery in a substantially dry,
`
`partially hydrated or hydrated ("wet") state. For example, a prosthetic implant utilizing a
`
`prepared tissue described herein may be packaged for delivery as a hydrated prosthetic implant.
`
`25 Accordingly, while a portion of the tissue preparation process may include drying the tissue so
`
`that it may be manipulated more easily, the tissue may then be hydrated at a later point in time
`
`prior to implantation, and it may be maintained in a hydrated condition up to and including
`
`packaging, delivery and implantation into a patient. Hydration of the tissue membrane portion
`
`occurs rapidly and begins with simple preparatory flushing of the tissue. Those skilled in the art
`
`30 will appreciate that one or more embodiments described herein provide a tissue 400 suitable for
`
`implanting in a human, wherein the implantable tissue may be allowed to dry prior to implanting
`
`and effectively rehydrated at the time of implanting, such as by flushing of the tissue at the time
`
`of implanting using saline or water.
`
`All embodiments described herein are described for use in human patients. However, all
`
`35
`
`embodiments described herein have application for use in veterinary medicine, such as equine
`
`13
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1554 of 2218
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`

`

`WO 2011/109433
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`medicine.
`
`PCT/US2011/026741
`
`The present invention may be embodied in other specific forms without departing from
`
`its spirit or essential characteristics. The described embodiments are to be considered in all
`
`respects only as illustrative and not restrictive. The scope of the invention is, therefore,
`
`5
`
`indicated by the appended claims rather than by the foregoing description. All changes which
`
`come within the meaning and range of equivalency of the claims are to be embraced within their
`
`scope.
`
`The one or more present inventions, in various embodiments, include components,
`
`methods, processes, systems and/or apparatuses substantially as depicted and described herein,
`
`10
`
`including various embodiments, subcombinations, and subsets thereof. Those of skill in the art
`
`will understand how to make and use the present invention after understanding the present
`
`disclosure.
`
`The present invention, in various embodiments, includes providing devices and
`
`processes in the absence of items not depicted and/or described herein or in various
`
`15
`
`embodiments hereof, including in the absence of such items as may have been used in previous
`
`devices or processes (e.g., for improving performance, achieving ease and/or reducing cost of
`
`implementation).
`
`The foregoing discussion of the invention has been presented for purposes of illustration
`
`and description. The foregoing is not intended to limit the invention to the form or forms
`
`20
`
`disclosed herein. In the foregoing Detailed Description for example, various features of the
`
`invention are grouped together in one or more embodiments for the purpose of streamlining the
`
`disclosure. This method of disclosure is not to be interpreted as reflecting an intention that the
`
`claimed invention requires more features than are expressly recited in each claim. Rather, as the
`
`following claims reflect, inventive aspects lie in less than all features of a single foregoing
`
`25
`
`disclosed embodiment. Thus, the following claims are hereby incorporated into this Detailed
`
`Description, with each claim standing on its own as a separate preferred embodiment of the
`
`invention.
`
`Moreover, though the description of the invention has included descriptions of one or
`
`more embodiments and certain variations and modifications, other variations and modifications
`
`30
`
`are within the scope of the invention (e.g., as may be within the skill and knowledge of those in
`
`the art, after understanding the present disclosure). It is intended to obtain rights which include
`
`alternative embodiments to the extent permitted, including alternate, interchangeable and/or
`
`equivalent structures, functions, ranges or acts to those claimed, whether or not such alternate,
`
`interchangeable and/or equivalent structures, functions, ranges or acts are disclosed herein, and
`
`35 without intending to publicly dedicate any patentable subject matter.
`
`14
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1555 of 2218
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`

`WO 2011/109433
`
`What is claimed is:
`
`CLAIMS
`
`PCT/US2011/026741
`
`1.
`
`A prepared tissue for medical use, comprising:
`
`a section of treated tissue harvested from a mammalian organism, the section of treated
`
`5
`
`tissue including an ultimate tensile strength of greater than about 15 MegaPascals.
`
`2.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue has a
`
`thickness of between about 50 to 500 micrometers.
`
`3.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue comprises a
`
`water content ofless than about 60% by weight of the section of treated tissue.
`
`10
`
`4.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue comprises a
`
`water content ofless than about 50% by weight of the section of treated tissue.
`
`5.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue comprises a
`
`water content of less than about 40% by weight of the section of treated tissue.
`
`6.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue is attached to
`
`15
`
`a frame ex vivo for at least one of: (a) surgical use; or (b) percutaneous implantation.
`
`7.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue does not
`
`include a matrix that has been exposed to a polymer infiltrate.
`
`8.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue is unbraided
`
`and uncompounded.
`
`20
`
`9.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue comprises an
`
`ultimate tensile strength of greater than about 25 MegaPascals.
`
`10.
`
`The prepared tissue of Claim 9, wherein the section of treated tissue is unbraided
`
`and uncompounded.
`
`11.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue has been
`
`25
`
`exposed to isopropyl alcohol before contacting the section of treated tissue with either
`
`glutaraldehyde or formalin.
`
`12.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue has been
`
`exposed to a solution containing formalin after pretreatment with isopropyl alcohol.
`
`13.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue has been
`
`30
`
`exposed to a solution containing glutaraldehyde after pretreatment with isopropyl alcohol.
`
`14.
`
`The prepared tissue of Claim 1, wherein the section of treated tissue comprises a
`
`pericardium tissue.
`
`15.
`
`A prepared tissue for medical use with a patient, comprising:
`
`a section of tissue harvested from a mammalian organism, wherein the section of tissue
`
`35
`
`is prepared ex vivo for future grafting or implantation in the patient, the section of tissue
`
`15
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1556 of 2218
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`

`

`WO 2011/109433
`
`PCT/US2011/026741
`
`including a thickness of about 50 to 500 micrometers and an ultimate tensile strength of greater
`
`than about 25 MegaPascals.
`
`16.
`
`The prepared tissue of Claim 15, wherein the section of tissue is unbraided and
`
`uncompounded.
`
`5
`
`17.
`
`The prepared tissue of Claim 15, wherein the section of tissue comprises a water
`
`content of less than about 40% by weight of the section of tissue.
`
`18.
`
`The prepared tissue of Claim 15, wherein the section of tissue is attached to a
`
`frame ex vivo for at least one of: (a) surgical use; or (b) percutaneous implantation in the patient.
`
`19.
`
`The prepared tissue of Claim 15, wherein the section of tissue does not include a
`
`10 matrix that has been exposed to a polymer infiltrate.
`
`20.
`
`The prepared tissue of Claim 15, wherein the section of tissue comprises a treated
`
`pericardium tissue.
`
`21.
`
`A method of preparing a tissue for medical use, comprising:
`
`providing a section of tissue harvested from a mammalian organism; and
`
`15
`
`causing osmotic shocking of the section of tissue by performing multiple rinses of the
`
`section of tissue with distilled water.
`
`22.
`
`The method of Claim 21, further comprising hydrating the section of tissue
`
`during a plurality of time intervals using distilled water.
`
`23.
`
`The method of Claim 22, further comprising not using saline for causing at least
`
`20
`
`one of the osmotic shocking and the hydrating of the section of tissue.
`
`24.
`
`The method of Claim 21, further comprising pretreating the section of tissue with
`
`glycerol before contacting the section of tissue with one or more of isopropyl alcohol,
`
`glutaraldehyde and formalin.
`
`25.
`
`The method of Claim 24, further comprising contacting the section of tissue with
`
`25
`
`a solution containing formalin after pretreating the section of tissue with glycerol.
`
`26.
`
`The method of Claim 24, further comprising contacting the section of tissue with
`
`a solution containing glutaraldehyde after pretreating the section of tissue with glycerol.
`
`27.
`
`The method of Claim 21, further comprising pretreating the section of tissue with
`
`isopropyl alcohol before contacting the section of tissue with either glutaraldehyde or formalin.
`
`30
`
`28.
`
`The method of Claim 27, further comprising contacting the section of tissue with
`
`a solution containing formalin after pretreating the section of tissue with isopropyl alcohol.
`
`29.
`
`The method of Claim 27, further comprising contacting the section of tissue with
`
`a solution containing glutaraldehyde after pretreating the section of tissue with isopropyl
`
`alcohol.
`
`16
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1557 of 2218
`
`

`

`WO 2011/109433
`
`PCT/US2011/026741
`
`30.
`
`The method of Claim 21, further comprising exposing the section of tissue to
`
`light energy for a period of time, the period of time extending until there is no further visible
`
`separation of lipid droplets from an exposed surface of the section of tissue.
`
`31.
`
`The method of Claim 30, wherein the light energy is at least equivalent to
`
`5
`
`exposing the section of tissue to a 50 watt incandescent light source with a flat radiant face
`
`situated at a distance of about 10 centimeters from the exposed surface for about 15 minutes.
`
`32.
`
`The method of Claim 21, wherein the section of tissue comprises a treated
`
`pericardium tissue.
`
`10
`
`15
`
`33.
`
`( a)
`
`A method of preparing a section of tissue for medical use, comprising:
`
`cleaning and decellularizing the section of tissue by performing multiple rinses of
`
`the section of tissue with distilled water;
`
`(b)
`
`rinsing the section of tissue with isopropyl alcohol for a first period of time of not
`
`less than about 7 days; and
`
`( c)
`
`contacting the section of tissue with one of
`
`(i)
`
`(ii)
`
`a formalin solution, or
`
`a glutaraldehyde solution
`
`for a second period of time of not less than about 6 days;
`
`wherein step (b) occurs sometime after step (a), and wherein step ( c) occurs sometime
`
`after step (b ).
`
`20
`
`34.
`
`The method of Claim 33, wherein for step (c):
`
`if the formalin solution is used, then the formalin solution comprises a concentration of
`
`about 1 - 37.5% formalin; and
`
`if the glutaraldehyde solution is used, then the glutaraldehyde solution comprises a
`
`concentration of about 0.1 - 25% glutaraldehyde.
`
`25
`
`35.
`
`The method of Claim 33, further comprising exposing the section of tissue to
`
`light energy for an exposure duration, the exposure duration extending until there is no further
`
`visible separation of lipid droplets from an exposed surface of the section of tissue.
`
`36.
`
`The method of Claim 35, wherein the light energy is at least equivalent to
`
`exposing the section of tissue to a 50 watt incandescent light source with a flat radiant face
`
`30
`
`situated at a distance of about 10 centimeters from the exposed surface for about 15 minutes.
`
`37.
`
`( d)
`
`The method of Claim 33, further comprising:
`
`rinsing the section of tissue with distilled water and isopropyl alcohol for a post-
`
`fixation period of time of not less than about 7 days;
`
`wherein step ( d) occurs sometime after step ( c ).
`
`17
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1558 of 2218
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`

`

`WO 2011/109433
`
`PCT/US2011/026741
`
`38.
`
`The method of Claim 33, wherein the section of tissue comprises an ultimate
`
`tensile strength of greater than about 25 MegaPascals.
`
`39.
`
`The method of Claim 33, wherein the section of tissue comprises a treated
`
`pericardium tissue.
`
`40.
`
`(a)
`
`(b)
`
`A method of preparing a section of tissue for medical use, comprising:
`
`contacting the section of tissue with distilled water;
`
`contacting the section of tissue with isopropyl alcohol for a pre-fixation period of
`
`time of not less than about 3 days; and
`
`( c)
`
`contacting the section of tissue with one of
`
`(i)
`
`(ii)
`
`a formalin solution, or
`
`a glutaraldehyde solution
`
`5
`
`10
`
`for a fixation period of time of not less than about 3 days; and
`
`(d)
`
`contacting the section of tissue with isopropyl alcohol for a post-fixation period
`
`of time of not less than about 3 days;
`
`15
`
`wherein step (b) occurs sometime after step (a), wherein step ( c) occurs sometime after
`
`step (b), and wherein step (d) occurs sometime after step (c).
`
`41.
`
`The method of Claim 40, wherein for step ( c):
`
`if the formalin solution is used, then the formalin solution comprises a concentration of
`
`about 1 - 37.5% formalin; and
`
`20
`
`if the glutaraldehyde solution is used, then the glutaraldehyde solution comprises a
`
`concentration of about 0.1 - 25% glutaraldehyde.
`
`42.
`
`The method of Claim 40, wherein for step (c):
`
`if the formalin solution is used, then the formalin solution comprises a concentration of
`
`about 8-12% formalin; and
`
`25
`
`if the glutaraldehyde solution is used, then the glutaraldehyde solution comprises a
`
`concentration of about 0.1-0.5% glutaraldehyde.
`
`43.
`
`The method of Claim 40, wherein the section of tissue comprises a treated
`
`pericardium tissue.
`
`18
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1559 of 2218
`
`

`

`WO 2011/109433
`
`PCT/US2011/026741
`
`1/5
`
`~100
`
`TISSUE PREPARATION
`
`r - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -~
`I
`I
`I
`I .,,..,,.
`1
`1
`I
`I
`
`~--------------- 1 ---------------- I
`
`DRY!NG
`
`, /
`
`END
`
`200
`
`.,,..,,. 300
`
`FIG. 1
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1560 of 2218
`
`

`

`WO 2011/109433
`
`PCT/US2011/026741
`
`2/5
`
`PERICARDIUM TISSUE HARVESTED
`
`204
`
`208
`
`TISSUE IS CLEANED AND DECELLULARIZED BY
`RINSING AND SEPARATING TISSUE WITH
`DISTILLED WATER
`
`200
`
`,,,✓ 220
`
`r - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 7 ,,,,,,
`(
`,,,
`:
`I I,.,.,,,
`
`TiONAL ADDITIONAL REMOVAL OF LIPIDS
`OP
`1
`100% GLYCEROL PRETREATMENT
`:
`I
`______________ T ______________ I
`
`r'f
`l _ _ _ _ _ _ _ _ _ _ _ _ _ _ t ______________
`I
`: ,.,.,,,,,,228
`r'i
`:
`LIGHT ENERGY EXPOSURE
`L--------------- J ______________ J
`I _____________________________ I
`
`224
`
`I
`I
`
`I
`
`1
`
`SECONDARY CLEANING
`
`RINSING VVITH DISTILLED \NATER
`
`RINSING WITH 25% ISOPROPYL ALCOHOL
`
`232
`
`236
`
`240
`
`244
`
`RINSING \NITH DISTILLED \NATER
`
`FIG. 2A
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1561 of 2218
`
`

`

`WO 2011/109433
`
`PCT/US2011/026741
`
`3/5
`
`0
`1
`
`~248
`
`FlXATION FOR COLLAGEN CROSS-LINKING
`(ONE OF THE FOLLOVV!NG)
`
`IMMERSION IN 1-37 .5% FORrv1AUN
`
`OR
`
`1Ml\•1ERSION lN 100% GLYCEROL
`
`OR
`
`IMMERSION lN 0:1 - 25% GLUTARALDEHYDE
`
`200
`
`v--
`---r--- 248 a
`
`248 b
`
`248 C
`
`IMMERSION IN 0.1-25'% GLUTARALDEHYDE (FILTERED --L-------248
`--L----- 24
`
`OR
`
`TO UMIT OL!GOMERIC CONTENT)
`
`OR
`
`IMMERSION IN ONE OF 248a, 248c, or 248d
`\NITH AMINO ACID ENRICHMENT
`
`ALCOHOL POST-FIXATION TREATMENT:
`
`RINSING IN DISTILLED WATER
`
`25%!SOPROPYLALCOHOL
`
`I
`
`I
`
`RINSING WITH DISTILLED WATER
`
`i
`•
`•
`
`---
`
`~
`
`d
`
`Be
`
`~252
`
`L-------256
`
`L-,-- 260
`
`r
`-----
`r ---
`
`~264
`
`~
`
`FIG. 28
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1562 of 2218
`
`

`

`WO 2011/109433
`
`PCT/US2011/026741
`
`4/5
`
`TISSUE IS DRIED
`
`TISSUE IS PLACED ON FLAT SURFACE
`
`304
`
`300
`
`V
`
`8
`/ 3 0
`
`V
`
`3 12
`TENSION AND/OR COMPRESSION LOAD ISv ~
`APPLIED TO TISSUE
`
`y
`
`TISSUE IS DRIED IN DARK CONDITIONS
`
`314
`~
`
`V
`
`FIG. 3
`
`, ,
`TISSUE INSPECTION
`AND
`ORIENTATION OF FIBER DIRECTION
`
`316
`
`V
`
`, ,
`---------------- ---------------- I
`I
`1--.,,.
`OPTIONAL SIZING OF TISSUE SHEET
`I
`I
`
`.,,. 320
`
`.,,..,,.
`
`TISSUE
`
`~ ,
`400
`
`F!G.4
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1563 of 2218
`
`

`

`WO 2011/109433
`
`PCT/US2011/026741
`
`Stress-Strain Curves of Wet Tissue
`50 r----,-----,-..;.......--r-;.._,.;.;......--y, _ _ _ -r-_ _ ---r' _ _ ___,..
`
`30
`
`if)
`if)
`w
`0:::
`~ to
`
`0
`
`~10.___ _ _ _ _ _ _ ....._ _ _ ......._ _ _ ___. ......... _ _ ..J.-_ _ --1 _ _ ~
`-0.2
`0
`0:3
`R1
`0.6
`-tl7
`0.4
`
`STRAIN
`Stress-strain curves in wet or hydrated state of five samples. Each curve corresponds
`to a separate sample.
`
`FIG. 5
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1564 of 2218
`
`

`

`(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`
`(19) \-Vorld Intellectual Property Organization
`International Bureau
`
`(43) International Publication Date
`9 September 2011 (09.09.2011)
`
`PCT
`
`1111111111111111 IIIIII IIIII 111111111111111 II Ill lllll 111111111111111 IIIII IIII IIIIIII IIII IIII IIII
`
`(10) International Publication Number
`WO 2011/109450 A2
`
`(51) International Patent Classification: Not classified
`
`(21) International Application Number:
`PCT/US201 li026763
`
`(22) International Filing Date:
`
`(25) Filing Language:
`
`(26) Publication Language:
`
`1 March2011 (01.03.2011)
`
`English
`
`English
`
`(30) Priority Data:
`61/309,109
`13/038,361
`
`us
`us
`(71) Applicant (for all designated States except US): COLIB(cid:173)
`RI HEART VALVE LLC [US/US]; 2150 W. 6th Ave,
`Suite M, Broomfield, CO 80020 (US).
`
`1 March 2010 (01.03.2010)
`1 March2011 (01.03.2011)
`
`(72) Inventors; and
`(75) Inventors/Applicants (for US only): FISH, R., David
`[US/US]; 6349 Vanderbilt Street, Houston, TX 77005
`
`(US). PANIAGUA, David [CR/US]; 3813 Dnunmond
`Street, Houston, TX 77025 (US).
`
`(74) Agent: YASKANIN, Mark, L.; Holme Roberts & Owen
`LLP, 1700 Lincoh1 Street, Suite 4100, Denver, CO 80203
`(US).
`
`(81) Designated States (unless otherwise indicated, for every
`kind of national protection available): AE, AG, AL, AM,
`AO, AT, AU, AZ, BA, BB, BG, BH, BR, BW, BY, BZ,
`CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO,
`DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT,
`HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP,
`KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, M.A, MD,
`ME, MG, MK, MN, J'vIW, MX, MY, MZ, NA, NG, NI,
`NO, NZ, OM, PE, PG, PH, PL, PT, RO, RS, RU, SC, SD,
`SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR,
`TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW.
`
`(84)
`
`Designated States (unless otherwise indicated, for eve1y
`kind o_f regional protection available): ARIPO (BW, GH,
`GM, KE, LR, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG,
`ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ,
`
`[Continued on next page]
`
`-
`;;;;;;;;;;;;;;;
`
`(54) Title: PERCUTANEOUSLY DELIVERABLE HEART VAL VE AND METHODS ASSOCIATED THEREWITH
`
`AA
`
`(57) Abstract: A prosthetic heart valve implantable by catheter
`without surgery includes a substantially "dry" membrane or tissue
`material. In at least one embodiment, the tissue is folded in a dry
`state to form a tissue leaflet assembly that is then attached to a
`frame to form an implantable prosthetic heart valve. Alternatively,
`one or more tissue leaflets are operatively associated with a frame
`to form an implantable prosthetic heart valve. The implantable
`prosthetic heart valve is subsequently pre-mounted on an integrat(cid:173)
`ed catlieter delivery system. The catheter delivery system that in(cid:173)
`cludes the implantable prosthetic heart valve is then packaged and
`transported while the tissue remains dry. The implantable pros(cid:173)
`thetic heart valve, while remaining substantially dry, can then be
`implanted into the receiving patient.
`
`;;;;;;;;;;;;;;; -
`-;;;;;;;;;;;;;;;
`;;;;;;;;;;;;;;; -
`
`;;;;;;;;;;;;;;;
`
`;;;;;;;;;;;;;;;
`
`1300
`
`1000
`
`-;
`
`;;;;;;;;;;;;;;
`;;;;;;;;;;;;;;;
`;;;;;;;;;;;;;;;
`
`FIG.13
`
`Edwards Lifesciences Corporation, et al. Exhibit 1003, p. 1565 of 2218
`
`

`

`WO 2011/109450 A2 1111111111111111 IIIIII IIIII 111111111111111 II Ill lllll 111111111111111 IIIII IIII IIIIIII IIII IIII IIII
`
`TM), E