PROCOAGULANT FACTORS SUITABLE FOR SUBSEQUENT AUTOLOGOUS USE

Abstract

Provided is a blood collection unit suitable for accelerated blood coagulation of whole blood for subsequent autologous or allogeneic use. The blood collection unit includes an inside surface that an activation site accelerating coagulation by having a high roughness. Further provided is a blood collection unit including an inside surface that has an activation site having a high roughness area, and an interior of the blood collection unit has been prepared with a pressure of no more than 255 mBar, and preferably a pressure of no more than 130 mBar.

Claims

1-12. (canceled)

13. A blood collection unit suitable for accelerated blood coagulation of whole blood for subsequent autologous or allogeneic use, comprising: an outer wall comprising a closed bottom end, an open top end, a side wall spanning between the ends, and a stopper for insertion into the open end to releasably seal the blood collection container, the stopper having a puncturable self-sealing septum or valve, an inner volume at least partially bounded by an inside surface of the outer wall, and a procoagulant environment, wherein the inner volume is prepared with a pressure of no more than 131.72 m Bar.

14. The blood collection unit according to claim 13, the blood collection unit further comprising: an activation site formed on the inside surface or formed on an insert in the inner volume of the blood collection unit, where the activation site has a roughness, R.sub.RMS, of at least 0.012 ?m.

15. The blood collection unit according to claim 14, wherein the activation site has a roughness, R.sub.RMS, of at least 0.4 ?m.

16. The blood collection unit according to claim 14, wherein the activation site is hydrophobic.

17. The blood collection unit according to claim 13, where the inside surface is hydrophobic.

18. The blood collection unit according to claim 13, wherein the blood collection unit is made of a hydrophobic material.

19. The blood collection unit according to claim 13, wherein the inside surface has not been corona treated.

20. The blood collection unit according to claim 13, wherein the inside surface has been corona treated.

21. The blood collection unit according to claim 13, wherein the blood collection unit is a tube or a double-ended needle.

22. The blood collection container according to claim 13, wherein the inner volume is prepared with a pressure of no more than 100 mBar.

23. The blood collection container according to claim 13, wherein the inner volume is prepared with a pressure of no more than 81.06 mBar.

24. The blood collection container according to claim 13, wherein the inner volume is prepared with a pressure of no more than 50.66 mBar.

25. The blood collection container according to claim 13, wherein the inner volume is prepared with a pressure of no more than 20.27 mBar.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0087] In the following, example embodiments are described according to the invention, where

[0088] FIG. 1 is a side view of the evacuated blood collection container means according to the present invention;

[0089] FIG. 2 is a view of the evacuated blood collection container means inserted into a first vacuum-sealed container means according to the present invention;

[0090] FIG. 3 is a view of the evacuated blood collection container means inserted into a first vacuum sealed container means inserted into a second vacuum sealed container means according to the present invention;

[0091] FIG. 4 is a side view of the blood collection container means while drawing blood through a double-ended needle.

[0092] FIG. 5 details measurement data demonstrating the effects of degrees of vacuum on coagulation of blood.

[0093] FIG. 6 illustrates various blood coagulation containers with roughened inside surfaces.

[0094] FIG. 7 illustrates readings during a controlled experiment of a rough inside surface.

[0095] FIG. 8 illustrates the effect of combining high vacuum and rough surface.

DETAILED DESCRIPTION OF THE DISCLOSURE

[0096] In the following, the invention is described in detail through embodiments thereof that should not be thought of as limiting to the scope of the invention.

[0097] FIG. 1 describes a side view of a blood collection container 100 comprising a bottom wall 102 and a side wall 103 forming a container with an open end and a closed end. A stopper 104 is fitted in said open end, said bottom wall 102, side wall 103 and stopper 104 enclosing an inner volume 101 of said blood collection container 100, said stopper comprising at least a mechanism for securely sealing said open end and maintaining a pressure differential, in a preferred embodiment making the blood collection container at least transitorily vacuum durable. Said stopper 104 further comprises a self-sealing puncturable septum 105 or a valve allowing the emptying of air from the inner volume 105 through any convenient method, such as a vacuum pump, said self-sealing puncturable septum 105 or valve further allowing the insertion of a needle or other tube-end without simultaneously allowing atmospheric air into the inner volume 101. This allows passage of blood through a needle into the blood collection container while maintaining a high vacuum in the inner volume 101.

[0098] Further depicted in FIG. 1 is an inner surface of said bottom wall 106, an inner surface of said side wall 107, and an inner surface of said stopper 108, said inner surfaces all facing and being in contact with said inner volume 101. These or some of these surfaces may be plasma-treated and/or abraded to accelerate coagulation speed further.

[0099] The blood collection container is prepared for use by inserting the stopper into its designated position in the open end of the blood collection container 100 and emptying the blood collection container of air through the self-sealing puncturable septum 105 or valve, said emptying of air further securing said stopper in its position by exerting a downward pressure due to the pressure differential over said stopper.

[0100] FIG. 2 is a side view of a blood collection container 100 inside a vacuum-sealed first container means 200, said first container means being at least transitorily vacuum durable, said blood collection container comprising a bottom wall 102, a side wall 103 defining a container with an open and a closed end, and a stopper 104 in said open end, said stopper further comprising a puncturable septum 105 or a valve for emptying the enclosed inner volume 101 of air thus creating a high vacuum, said puncturable septum or valve further allowing drawing blood into the blood collection container without simultaneously allowing atmospheric air into the inner volume 101.

[0101] The blood collection container is inserted into the first container means 200 which is emptied of air thus ensuring that it encloses said blood collection container tightly, whereupon said first container means is vacuum-sealed thus lowering the pressure differential over the bottom wall, side wall, and stopper of said blood collection container thus improving the effective vacuum durability of said blood collection container. Any convenient sealing method may be used and in one embodiment, when a convenient vacuum has been achieved inside the first container means, it is heat-sealed at the sealing line 201.

[0102] FIG. 3 is a side view of a vacuum-sealed second container means 300 being substantially vacuum-durable, said second container means comprising a first container means 200 further comprising a blood collection container prepared with a high vacuum.

[0103] The three containers taken together, i.e. the blood collection container, the first container means 200, and the second container means 300, make up a blood collection kit of parts prepared for transportation, storage and handling.

[0104] Preferably, a first container means 200 vacuum-sealed and comprising a blood collection container is sterilised, whereupon it is inserted into a second container means 300, said second container means then being emptied of air thus ensuring that it encloses said first container means tightly, whereupon said second container means is vacuum-sealed thus lowering the pressure differential over the enclosed vacuum barriers and improving the effective vacuum durability of said blood collection container. In one embodiment, when a convenient vacuum has been achieved inside the second container means such as a high vacuum, it is heat sealed at the sealing line 301.

[0105] FIG. 4 is a side view of a blood collection kit of parts 4000, the kit of parts comprising a blood collection container 100 and a double-ended needle 400 after drawing blood from a patient and disassembling the kit of parts for further processing of the blood.

[0106] The blood collection container 100 comprises a bottom wall 102, a side wall 103 defining a container with an open and a closed end, and a stopper in said open end, said stopper further comprising a puncturable septum 105 or a valve for emptying the enclosed inner volume 101 of air thus creating a high vacuum, said puncturable septum or valve further allowing drawing blood into the blood collection container without simultaneously allowing atmospheric air into the inner volume 101. The blood collection container further comprises a blood sample 405.

[0107] The double-ended needle comprises a first needle end 401, a second needle end 402, an intermediate tube 403, and an inner volume 404 running the axial length of said double-ended needle, the inner volume allowing the transport of blood from the patient to the blood collection container. In one embodiment of the invention, the inner volume further comprises coagulation accelerating objects, such as one or more glass beads.

[0108] In another embodiment of the invention, the double-ended needle further comprises a membrane or pressure valve controlling the flow of blood after injecting the first needle end into a blood-filled cavity such as the vein of a patient and prior to inserting the second needle-end into the blood collection container.

[0109] In one embodiment of the invention, the blood collection kit of parts further comprises a first container means 200 and a second container means 300.

[0110] A high vacuum may be introduced into the blood collection container by any convenient method. Preferably a blood collection kit of parts 4000 is unpackaged thus allowing access to the blood collection container 100 from within a first and a second container means, the blood collection container being previously prepared with a high vacuum. Then the first needle head 401 enters a convenient volume of the patient, typically a vein, whereupon blood begins running through the inner volume 404 of the double ended needle and simultaneously blocking fluid communication from the blood collection container to outside ambient pressure through the double-ended needle at the first needle end 401. In another embodiment, a membrane or pressure valve at a convenient location inside the double-ended needle, preferably close to the second needle-end, blocks the flow of blood prior to bringing the two inner volumes 101, 404 into fluid communication. This allows easier handling by not needing as careful handling and observation to ensure the blood does not run out of the second needle end prior to puncturing the self-sealing septum as well as a controlled and limited depressurisation as consequence of bringing the two inner volumes into fluid communication since the blood will have pushed a certain controlled, preferably high amount of the air out of the double-ended needle. For example, this membrane or valve comprises rubber or another convenient material. Bringing the inner volumes into fluid communication breaks or opens the membrane or opens the valve and allows the blood to flow.

[0111] In the case of using a double-ended needle with a membrane, valve or without either, a second needle end 402 then punctures the self-sealing septum of a blood collection container 100 which brings the high vacuum inner volume of the blood collection container 101 into fluid communication with the inner volume of the double-ended needle 404, this fluid communication decreasing the pressure in the inner volume of the double-ended needle 404 and so actuates an accelerated blood draw.

[0112] FIG. 5 is measurement data of the various effects on coagulation speed by using 90% (101 mBar), 92% (81 mBar) and 98% (20 mBar) vacuum in a blood collection container during blood filling, Thereafter inserted into a centrifuge, where the coagulation process is observed by way of the translucency of the top part of the blood sample.

[0113] In a first phase, translucency increases continually as the blood separates and blood plasma remains in the uppermost part due to it comprising elements relative to the rest of the blood lower Svedberg values, which means forces like gravity, and the centripetal force acts less strongly on these elements and may be thought of as an alternative property to mass for the sake of this invention.

[0114] In a second phase, fibrin in the blood plasma begins polymerising thus reducing translucency as the blood plasma becomes more opaque. This reduces transmission counts. Table 1 describes the events shown in FIG. 5:

TABLE-US-00001 TABLE 1 90% 92% 98% peak transmission count at 7.94 minutes 7.45 minutes 6.91 minutes post-peak lowest transmission 9.93 minutes 8.74 minutes 7.62 minutes count at

[0115] FIG. 6 illustrates various embodiments of the invention. All embodiments shown in FIG. 6 illustrate a blood collection container as previously described with a bottom wall 102, an open end with a stopper 104, and a sidewall spanning between 103. The stopper has a self-sealing septum 105.

[0116] The inside surface of the blood collection container 100 is provided with an activation site 600. In FIG. 6a, the bottom surface is provided with an activation site 600. The activation site has a high roughness average (R.sub.a) which has been found to increase coagulation speed significantly. By providing the activation site 110 at the bottom of the blood collection container 100, the blood will be pressed against it during centrifugation thus enhancing the effect hereof.

[0117] FIG. 6b illustrates an embodiment, where the activation site 600 is provided on the sidewall. The embodiments shown in FIGS. 6a and 6b may be combined to provide more activation sites and/or spreading these around in the blood collection container 100.

[0118] FIG. 6c illustrates an embodiment of the invention, where the activation site 600 is provided in an insert 602. The illustrated insert is a float which has an inner volume with a lower density than blood thus allowing lifting the float during or after centrifugation. Thereby, as the blood travels through the central hole 604 to beneath the float 602 during lifting the float 602, the blood will move over the activation site 600 on the insert 602 and thus expose more blood to the activation site 602. There may be provided any kind of channels through or next to the insert in accordance with the invention.

[0119] FIG. 6d illustrates a double-ended needle 400 mostly as previously described. The double-ended needle 400 has on its inner surface 406 an activation site 600 that is an area with a high roughness. As the blood passes the activation site 600, the coagulation cascade is initiated. This allows early and thorough coagulation activation.

[0120] Generally, the inner surface of the collection unit being the double-ended needle or the container should at least partly have a surface with a high roughness (activation site), whereby the surface is non-smooth, and where this non-smooth surface influences the coagulation speed. As mentioned above, the non-smooth surface could be provided on the sidewalls or bottom walls of the container itself or it could be provided on inserts positioned in the container.

[0121] The inner surface of a plastic container could e.g. be made non-smooth to increase the roughness by abrading the surface e.g. using abrading machinery or manually using abrading material.

[0122] FIG. 7 shows a controlled experiment of the effect of a rough surface on blood coagulation 710, even when the surface is of a hydrophobic material. The experiment measured optical transmittance 714 and RPM of the centrifuge 716 as functions of time 712.

[0123] The experiment was conducted on two identical samples. One was exposed to a rough inside surface, the test sample 720, 720 and the other were not exposed to a rough inside surface, i.e. the control sample 724, 724. Centrifuge work speed 728 was also measured. The variance between 720 and 724 on the one side and 720 and 724 on the other side is due to two sensor locations of the container during centrifugation.

[0124] The optical transmittance 714 changes during the experiment as a result of the coagulation. At first, when the blood is still mixed and in liquid form, the optical transmittance is low as light cannot penetrate the blood. During centrifugation, the blood is separated into several layersat first a plasma layer at the top being yellowish transparent, a buffy coat layer being a more opaque white in the middle, and a dark red layer of erythrocytes (red blood cells) at the bottom. These layers emerge as the heavier molecules are drawn to the bottom of the blood collection container. Subsequently, fibrin gradually polymerises from the plasma layer and descends onto the buffy coat layer. Without the fibrin, the top layer is then serum which is even more transparent than plasma. The polymerised fibrin is an opaque yellowish white.

[0125] The optical transmittance 714 is measured at the top of the container in what becomes the plasma fraction during centrifugation. Therefore, the optical transmittance increases steadily as heavy and generally light-absorbing molecules descend. This is initially comparative between the test sample 720 and the control sample 724. However, the translucency drops off for the test sample 720 earlier than for the control sample 724 thus indicating that fibrin is polymerising significantly earlier due to the rough surface.

[0126] FIG. 8 shows a controlled experiment of the effect of combining a high vacuum with a rough surface on blood coagulation 810. The experiment measured optical transmittance 814 as function of time 812. The blood considered has been exposed to centrifugation during the time considered. Thus, the optical transmittance 814 changes during the experiment as a result of the coagulation taking place during centrifugation. The process of coagulation and its impact on optical transmittance 814 is identical to the one described in FIG. 7.

[0127] The experiment of FIG. 8 was conducted on four identical whole blood samples. The samples were drawn into four different blood collection containers all comprising different environments: The first sample 831 was exposed to a vacuum of 80% (202.65 mBar), the second sample 832 was exposed to a vacuum of 98% (20.27 mBar), the third sample 833 was exposed to a vacuum of 80% (202.65 mBar) and a surface comprising an activation site having a high roughness, and the fourth sample 834 was exposed to a vacuum of 98% (20.27 mBar) and a surface comprising an activation site having a high roughness.

[0128] Considering first the samples exposed to vacuum only, the optical transmittance 814 of both the first 831 and the second 832 samples are seen to increase steadily and comparatively. However, the optical transmittance 814 drops off for the second sample 832 earlier than the first sample 831 thus indicating that fibrin is polymerising, i.e. coagulation is taking place significantly earlier in the high vacuum of the second sample 832 than for the first sample 831 comprising a relatively lower vacuum.

[0129] Considering the third 833 and the fourth 834 sample, both being exposed to an activation site having a high roughness, the optical transmittance 814 is seen to increase steadily and comparatively at first. However, it is seen that the fourth sample 834, which is further exposed to a relatively higher vacuum (98%, 20.27 mBar) than the third sample 833, drops off earlier than said third sample which is exposed to a relatively lower vacuum (80%, 202.65 mBar) than the fourth sample 834. Furthermore, it is seen that the fourth sample 834 coagulates earlier than the second sample 832 despite both being exposed to the same vacuum. The same is seen for the third sample 833 coagulating earlier than the first sample 831.

[0130] Thus, from the experiment 810, it is seen partly that a high vacuum causes faster coagulation of blood than a relatively lower vacuum and partly that a surface comprising an activation site having a high roughness causes coagulation faster than a surface not comprising such activation site. Most importantly, it is further seen that the combination of a high vacuum (98%, 20.27 mBar) and a surface comprising an activation site having a high roughness (i.e. the fourth sample 834) causes coagulation of blood before any other combinations (the first 831, the second 832, and the third 833 sample).

[0131] High Vacuum=A pressure of no more than 255 mBar, 253.31 mBar, 202.65 mBar, 151.99 mBar, 101.33 mBar, 91.19 mBar, 81.06 mBar, 70.93 mBar, 60.80 mBar, 50.66 mBar, 40.53 mBar, 30.40 mBar, 20.27 mBar, 10.13 mBar, 5.07 mBar, 1.01 mBar, 0.10 mBar or less, and where the preferable range of pressure is 0.10 mBar-101.33 mBar or less.

[0132] In another embodiment of the invention, said high vacuum corresponds to a vacuum of at least 75% (253.31 mBar), 76% (243.18 mBar), 77% (233.05 mBar), 78% (222.92 mBar), 79% (212.78 mBar), 80% (202.65 mBar), 81% (192.52 mBar), 82% (182.39 mBar), 83% (172.25 mBar), 84% (162.12 mBar), 85% (151.99 mBar), 86% (141.86 mBar), 87% (131.72 mBar), 88% (121.59 mBar), 89% (111.46 mBar), 90% (101.33 mBar), 91% (91.19 mBar), 92% (81.06 mBar), 93% (70.93 mBar), 94% (60.80 mBar), 95% (50.66 mBar), 96% (40.53 mBar), 97% (30.40 mBar), 98% (20.27 mBar), 99% (10.13 mBar), 99.5% (5.07 mBar), 99.9% (1.01 mBar) or even 99.99% (0.10 mBar) or above as measured linearly, where 0% vacuum is 1013.25 mBar and 100% vacuum is 0 mBar, and where the preferable range of vacuum is 90%-99.99% or above.

[0133] V.sub.r, is a relatively defined inner volume 404 of a double-ended needle 400=1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% or less relative to the volume of the blood collection container 100.

[0134] V.sub.a, is an absolutely defined inner volume of the double-ended needle, where it corresponds to V.sub.r for a blood collection container of 10 ml=0.1 ml being 1% of 10 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.5 ml, 0.6 ml, 0.7 ml, 0.8 ml, 9 ml or 1.0 ml being 10% of 10 ml. Vacuum Durability=a quality of a boundary enclosing an environment defining its ability to maintain a percentage of a pressure differential over said boundary over time, specifically where there is a pressure drop from the exterior to the enclosed environment. Two distinct degrees of vacuum durability is described; transitory vacuum durability and substantial vacuum durability, where:

[0135] A boundary with a transitory vacuum durability maintains at least 10% of its pressure differential over 24 hours, 48 hours, 72 hours, 96 hours, 120 hours, 144 hours, 168 hours or more, where the initial pressure inside the enclosed environment is a high vacuum and relative to 1013.25 mBar at ambient temperature outside.

[0136] A boundary with a substantial vacuum durability maintains at least 98% of its pressure differential over 1 week, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 12 weeks or more, where the initial pressure inside the enclosed environment is a high vacuum and relative to 1013.25 mBar at ambient temperature outside.

[0137] Blood collection container=a container specifically designed for storage and collection of blood immediately after drawing blood from a patient. Such blood collection containers may be used in subsequent treatment of the blood such as in producing LeucoPatch?, or it may only be used for transportation and storage of the blood for later use. Blood collection containers are most conveniently produced in glass, polyethylene terephthalate (PET) and/or polystyrene (PS).

[0138] Double-ended needle=typically a tube with a needle in both ends, preferably being able to puncture skin and a vein in the one end and a puncturable septum in the other end. In other embodiments of the invention, the double-ended needle may be regarded as any tube the ends of which are injected into an upstream blood-filled cavity, such as a vein or a blood container, and a downstream blood collection container despite the ends not being actual needles, the term double-ended needle encompassing catheters and tubes as well. Double-ended needles are conveniently produced in silicone, polyurethane, polyethylene and/or Teflon/PFTE.

A. A blood collection container (100) having a bottom wall (102) and a side wall (103) defining an open end, a stopper (104) in said open end, said stopper (104) further comprising a puncturable self-sealing septum (105) or valve, said elements defining an interior volume (101) of said blood collection container (100), characterised in that the interior volume (101) has been prepared with a pressure in its inner volume (101) of no more than 255 mBar.
B. A blood collection container (100) according to embodiment A, wherein said inner volume (101) is prepared with a pressure of no more than 200 mBar.
C. A blood collection container (100) according to any of embodiments A-B, wherein said inner volume (101) is prepared with a pressure of no more than 130 mBar.
D. A blood collection container (100) according to any of embodiments A-C capable of retaining at least 10% of the pressure differential between the high vacuum in its inner volume (101) relative to ambient temperature and pressure on the outside after 24 hours.
E. A blood collection container (100) according to any of embodiments A-D further comprising components adapted to counteract anti-coagulants added to the blood, such as calcium, inside the blood collection container (100).
F. A blood collection container (100) according to any of embodiments A-E, where at least part of the inner surfaces (106, 107, 108) of said blood collection container (100) surrounding the inner volume (101) are corona-treated.
G. A blood collection container (100) according to any of embodiments A-F further comprising a blood coagulation acceleration agent inside the blood collection container (100).
H. A blood collection container (100) according to any of embodiments A-G further comprising an activation site (600) formed in the inside surface (106, 107, 108), the activation site having a high roughness.
I. A blood collection container (100) according to embodiment H, where the activation site (600) has a roughness of at least 0.012 ?m.
J. A blood collection container (100) according to any of embodiments H-I, where said activation site is hydrophobic.
K. A blood collection container (100) according to any of embodiments H-J, where the inside surface (106, 107, 108) is hydrophobic.
L. A blood collection container (100) according to any of embodiments H-K made of a hydrophobic material.
M. A blood collection kit of parts (4000) comprising a blood collection container (100) for accelerated coagulation of blood according to any of embodiments A-L, wherein said kit of parts further comprises a first container means (200) and a second container means (300), said second container means (300) being capable of retaining at least 98% of the pressure differential between the vacuum in its inner volume relative to ambient temperature and pressure on the outside after one week, the second container means (300) vacuum being sealed around said first container means (200) which is capable of retaining at least 10% of the pressure differential over its enclosing walls after 24 hours, the first container means (200) being vacuum-sealed around said blood collection container (100).
N. A blood collection kit of parts (4000) according to embodiment M, where the first container means (200) comprises PS and/or PET and the second container means (300) comprises aluminium or aluminium foil.
O. A blood collection kit of parts (4000) comprising a blood collection container (100) for accelerated coagulation of blood according to any of embodiments A-N further comprising a double-ended needle (400) comprising a first needle end (401), a second needle end (402), an intermediate tube (403), and an inner volume (404) running the axial length of said double-ended needle (400), where the combination of the degree of vacuum prepared in said inner volume of the blood collection container (101) and said inner volume of said double-ended needle (404) is such that bringing the two inner volumes (101, 404) into fluid communication retains a pressure of no more than 255 mBar, preferably no more than 200 mBar, even more preferably no more than 130 mBar inside the combined inner volumes (101, 404).
P. A blood collection kit of parts (4000) according to embodiment 0, wherein said double-ended needle (400) further has a procoagulant environment (404).

[0139] A blood collection kit of parts (4000) according to embodiment P, wherein said procoagulant environment comprises an activation site (600) on an inner surface (406) of said double-ended needle (400) having a high roughness.