Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
  • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
  • G01N33/483—Physical analysis of biological material
  • G01N33/487—Physical analysis of biological material of liquid biological material
  • G01N33/49—Blood
  • G01N33/4905—Determining clotting time of blood
• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N11/00—Investigating flow properties of materials, e.g. viscosity, plasticity; Analysing materials by determining flow properties
  • G01N11/02—Investigating flow properties of materials, e.g. viscosity, plasticity; Analysing materials by determining flow properties by measuring flow of the material
  • G01N11/04—Investigating flow properties of materials, e.g. viscosity, plasticity; Analysing materials by determining flow properties by measuring flow of the material through a restricted passage, e.g. tube, aperture
  • G01N11/08—Investigating flow properties of materials, e.g. viscosity, plasticity; Analysing materials by determining flow properties by measuring flow of the material through a restricted passage, e.g. tube, aperture by measuring pressure required to produce a known flow

Definitions

• Device and method for detecting coagulation functions of global, especially primary, hemostasis are provided.
• the invention relates to a device according to the preamble of patent claim 1 and a method according to the preamble of patent claim 58.
• a volume flow of blood to be examined is transported from a storage vessel by means of a piston-cylinder arrangement through a flow path of a reaction device designed as an aperture. Aggregation and / or coagulation of blood components, in particular platelets, results in increasing blockage of the aperture.
• the pressure drop that forms on the outlet side of the aperture is detected and measured in a pressure measurement space that is free of the blood to be examined via a supply line to a pressure sensor.
• the pressure measuring chamber is located below a piston surface of the piston which moves vertically upward and delimits the working chamber of the piston-cylinder arrangement.
• the blood to be examined is transported upwards from a storage room located below the aperture.
• EP 0 635 720 AI discloses a method and a device in which the accumulation or aggregation of the blood platelets is initiated under certain flow conditions. Due to a rotational movement that the blood to be examined executes against a surface, shear forces occur on the surface. Platelets accumulate on the bottom of the container in which the blood to be examined is located. The deposited blood particles are then evaluated by electron microscope scanning or optical image analysis or the like.
• Platelets can be examined without unphysiological, mechanical or chemical influences.
• whole blood is drawn from a patient's vein using a cannula to which the reaction device is connected.
• the reaction device contains a collagen plate with an exactly drilled opening with a diameter of 0.5 mm.
• the blood drawn flows through this opening at a speed of 8 cm / s, each platelet passing through the opening in the collagen within approximately 50 ms.
• a constant flow is controlled by a mechanically NEN pump forced, which is connected to the reaction device.
• the blood is aspirated at a rate of 0.94 ml / min. As the platelets pass through the collagen opening, they are retained and increasingly close the borehole in the channel of the reaction device.
• the time in which the suction pressure rises from 50 to 150 mbar is determined. Furthermore, a device can be placed between the cannula and the reaction device, through which the solution of a substance is continuously supplied, the influence of which on the platelet function on collagen can be checked. The pressure is measured in the suction hose, which is filled with 0.9% NaCl solution.
• a measuring device is known from US Pat. No. 5,662,107 with which thrombus formation is measured in vitro under simulated in vivo conditions.
• blood is pumped at a constant flow rate through a channel which consists of a material which promotes thrombus formation or is coated with such a material.
• a pressure measurement is carried out upstream and downstream of the thrombus-forming unit and the pressure difference is evaluated as a tendency to form a thrombus.
• the importance of the shear rate for the attachment of platelets and the activation of coagulation is pointed out. However, this does not take into account the fact that as the addition of platelets increases, the shear rate increases uncontrollably if the flow rate is maintained.
• the object of the invention is to provide a device and a method of the type mentioned in the introduction, in which accurate and reproducible measurement results are achieved with little measurement effort. This object is achieved in the device by the characterizing features of one of claims 1, 2 and 3 and in the method by the characterizing features of claim 58.
• the device provides the prerequisite for the inexpensive design of a multi-channel machine for use in large laboratories as well as for a small device with e.g. a measuring channel, for use under difficult conditions in the area close to the patient (Point of Gare).
• the method enables the measurement sequence to be carried out under controlled shear conditions in the changing reaction opening and thus to form new important diagnostic approaches.
• Whole blood or platelet-rich plasma can be used to measure the coagulation functions of global, especially primary, hemostasis.
• Sodium citrate, hirudin and other substances can be used as anticoagulants.
• Activators such as e.g. Collagen, adenosine diphosphate, thrombin and other substances listed below are located on the boundary surfaces of the reaction openings or can be added to the blood sample before or during the measurement.
• the pressure measuring space is located below the storage space from which the blood to be examined is conveyed.
• the reaction device is located between the storage room and the pressure measuring room. After the blood has passed through the reaction device, which can be arranged below, for example in the floor area of the storage space, the blood to be examined is passed through the central flow opening past the pressure measuring space into a pressure-tight blood collection space where it collects during the measuring process.
• the pressure measuring chamber is located above the surface of the blood collected in the pressure-tight blood collection chamber.
• a pressure proportional to the delivery pressure or the same pressure as the delivery pressure prevails in the pressure measuring chamber.
• a piston-cylinder drive arrangement is preferably used to generate the delivery pressure.
• the pressure measuring space can be located in the working space of the piston-cylinder arrangement, in which the delivery pressure for the blood to be delivered through the reaction opening of the measuring device prevails.
• the pressure in the pressure measuring room then corresponds to the delivery pressure.
• a piston surface forms the boundary of the working space as the working surface.
• the pressure measuring space is thus formed by the cylinder of the piston-cylinder arrangement and one or more spacing means, in particular by part of the reaction device.
• the piston-cylinder arrangement is preferably located below a storage vessel in which the storage space for the blood to be examined is provided.
• the piston-cylinder arrangement can form the blood collection space with the expanding cylinder interior (work space).
• the device supplying the delivery pressure e.g. B. the piston-cylinder arrangement, however, as will be explained, can also be arranged outside the blood collection space.
• a pressure sensor can be arranged in it, which delivers corresponding signals.
• a pressure line for example a hollow needle, is used, which is passed through the wall of the pressure measuring space in a pressure-tight manner and is connected to a pressure measuring device e.g. B. pressure sensor, is connected outside the pressure measuring space.
• the hollow needle preferably has on the through the wall of the pressure measuring space passed end, a tip in the manner of an injection cannula.
• Pressure measuring chamber surrounding wall made of plastic so that it can be pierced with the needle to insert the needle end pressure-tight in the pressure measuring chamber.
• the storage vessel and the cylinder of the piston-cylinder arrangement can be formed from one piece.
• the reaction device via which the blood is led through the central flow opening past the pressure measuring space.
• the piston-cylinder arrangement located below can also be formed in a preferred manner by a blood withdrawal syringe in which there is anticoagulated blood drawn from the patient, the piston of which can be connected to the drive 17 via a coupling.
• a hollow needle which is attached with one end in a pressure-tight manner in the central flow opening, penetrates the sealing insert in the syringe adapter pointing upwards and thus connects the blood collection space (storage space) via the reaction device to the storage space (blood collection space) of the blood collection syringe.
• the blood supply space is formed in the working space of the cylinder of the withdrawal syringe.
• the flow opening of the hollow needle can be designed as a feed opening, which preferably acts as a shear opening.
• the plunger in the syringe cylinder is preferably moved upwards towards the reaction device and, with its working surface, pushes the blood column formed by the inner wall of the cylinder upwards when the vent valve 20 is open, until the upper blood level is at a precisely defined one Position is detected by a level sensor, which then signals the control unit, the reference position, from which the measurement can then start.
• the reference position defines the volume of the air cushion, which forms the blood-free pressure measurement space and is limited by the outer surface of the hollow needle, the blood level, the inner wall of the syringe adapter and the underside of the sealing insert 26.
• This embodiment is particularly suitable for use in the POC (Point of Care) area near the patient because the cumbersome pipetting of the blood sample is no longer necessary.
• a pressure line carrying the conveying pressure in particular gas pressure line
• gas pressure line can be provided, which is guided pressure-tight from the outside into the blood-free pressure measuring space through a container wall of the pressure-tight collecting space.
• the container wall is preferably made of plastic, in particular polyethylene, or another material that presses tightly against the outside of the pressure line when it is punctured.
• the pressure line can preferably be designed as a hollow needle (cannula) with a tapering needle end. The pointed end of the needle is pushed through the container wall into the pressure measuring chamber.
• the conveying device can be designed in a known manner as a suction / pressure pump, piston-cylinder arrangement or the like, which in conjunction with the drive generates the conveying pressure in the pressure line.
• the pressure line led through the container wall of the storage space or the collecting space carries both the delivery pressure and the pressure to be measured.
• the pressure line outside the container wall is connected to a pressure measuring device, in particular pressure sensor, via a branched pressure measuring line.
• the delivery pressure corresponds to the pressure to be measured and prevailing in the blood-free pressure measuring room.
• the reaction device is preferably arranged above the collecting space and with the collecting space in a common housing. Furthermore, the storage space above the reaction device can also be provided in the common housing. For this purpose, the storage space and at least parts of the reaction device and the collecting space for forming the common housing can consist of one piece. However, it is also possible to design the separate reaction device to be liquid-tight in the common housing.
• the storage space can also be formed by the cylinder interior of a blood sampling syringe, the cylinder interior being connectable to the reaction device via a hollow needle.
• the hollow needle can be designed as a cannula of the blood collection syringe containing the storage space, which can be guided pressure-tight from the outside with its needle tip to the reaction point of the reaction device.
• the carrier material of the reaction device can be designed such that it rests on the exterior of the hollow needle after it has pierced through the hollow needle.
• the hollow needle which leads to the reaction point of the reaction device, to the reaction device in a pressure-tight manner and to push its free needle end through a sealing wall of the storage space which can be fitted from above, for example the cylinder interior of a syringe.
• the sealing wall of the storage space also consists, as already explained, of a material that lies sealingly around the hollow needle.
• the hollow needle preferably forms the supply opening for the blood, from the storage space to the reaction opening and can also preferably serve as a shear opening. This embodiment is particularly suitable for use in the POC (Point of Care) area close to the patient because the cumbersome pipetting of the blood sample is no longer necessary.
• POC Point of Care
• An embodiment is preferably used for the detection of coagulation properties of global, in particular also primary, hemostasis, the blood to be examined being conveyed by a delivery device, in particular a piston-cylinder device, which can be driven by a drive device, from a storage room can be conveyed to the blood collection space via one or more reaction openings, which can also count as shear openings, in the reaction device.
• Feed openings which can simultaneously act as shear openings, can be arranged upstream or downstream of the reaction opening or openings, the surfaces of which are hydrophobic in order to avoid deposits of blood components there.
• the boundary surfaces of reaction openings can also be made hydrophobic or be provided with a roughness pattern.
• the boundary surface or partial surfaces thereof, one or more reaction openings or locations, at which blood components possibly attach or react under the action of shear forces, can be made, for example, of hydrophilic, optionally also hydrophobic plastic, glass, porous or non-porous bioactive foils / membranes, collagen membranes, porous Membranes (e.g. cellulose acetate) exist or are covered with them and can continue to be used in addition or if necessary for different questions of coagulation or plate formation.
• Chen reaction be designed reactive, in which they can be bioactive coated, soaked or coated, z. B.
• ADP adenosine philosophy
• adrenaline adrenaline
• fibronectin thrombospondin
• US 5,854,067; US 6,662,107 US 5,854,067; US 6,662,107
• the flow rate of the blood is proportional to the radius of the blood vessel and is lower on the wall of the vessel than in the center of the vessel.
• the difference in speed of adjacent layers of liquid flowing parallel to one another produces a shear effect between these layers. It is largest at the vascular wall and decreases towards the vascular center.
• the local shear rate which corresponds to the speed gradient between two adjacent flowing layers of liquid, influences the shear stress and is directly proportional to it. Accordingly, different shear rates prevail in different types of vessels on the surface of the vessel walls.
• Physiological shear rates are ⁇ 100 s "1 in large veins.
• the wall shear rates vary between 100 - 1000 s _1 and reach about 1500 s _1 in arterioles.” In the coronary arteries, the average shear rate is around 650 s _1 . Extremely high shear rates of approximately 3000 s _1 to a maximum of 40 000 s _1 exist in arteriosclerotic vessels, which vary depending on the amount of shear stress certain types of cells, in particular in the case of platelets, the external forms and responsiveness as well as the binding behavior of the membrane or plasma proteins. It is known that normal, but in particular activated blood platelets, with increasing increase in the shear rate, also adhere increasingly to, for example, collagen surfaces and then aggregate (arteriosclerosis).
• platelets with inhibited function can less and less adhere to, for example, collagen surfaces and thus aggregate (tendency to bleed) due to the action of ASA (acetylsalicylic acid) or an existing von Willebrand disease, with increasing shear rate.
• ASA acetylsalicylic acid
• This finding can be used in the invention for the sensitive diagnosis of platelet functions of primary hemostasis by controlling a blood volume flow through an aperture or reaction opening coated with, for example, collagen in such a way that there is a predetermined, in particular constant, shear rate due to the accumulation and aggregation of platelet reducing or occluding aperture / reaction opening is maintained.
• the volume flow of the blood to be examined through the aperture / reaction opening can also be set as a function of any predetermined shear rate or shear force characteristic.
• the delivery flow created by the controlled movement of the piston in the piston-cylinder arrangement generates a delivery pressure that builds up in accordance with the flow resistance in the reaction opening (s).
• the blood flow creates shear conditions or flow conditions in the reaction opening (s) with the cooperation of which platelets, depending on their functionality, are designed to adhere and aggregate at the boundary surfaces in the reaction opening (s) which are bioactive or capable of attachment and thereby reduce the open cross section of the reaction opening or this reaction opening by forming a thrombus entirely can close or there is a change in the flowability of the blood by the onset of global blood coagulation, in particular by the influence of the reactively designed boundary surface (s) of the reaction opening (s) or by the influence of supplied activators
• the shear rate or shear force which is effective at the reaction point or in the reaction opening, follows a predetermined characteristic curve, which preferably corresponds to a constant shear rate / force or another predetermined shear or flow rate curve.
• the measurement results and evaluation results obtained in this way correspond to the actual attachment and aggregation behavior of the platelets in accordance with the platelet reaction of the primary hemostasis or the coagulation behavior of the global hemostasis.
• the then existing volume flow and / or the volume flow rate, which has flowed through the reaction device after a certain predetermined measurement time, or the elapsed time and / or the volume flow rate when the volume flow has reached a predetermined value or against Zero goes (DE 35 41 057 AI).
• the elapsed time and / or the volume flow then present can be evaluated as platelet parameters.
• the measured pressure change and / or the volume flow rate reached after a predetermined time or the elapsed time when a predetermined pressure value and / or a volume flow rate is reached can be used as measurement parameters for the global, in particular primary, hemostasis.
• a further measurement evaluation takes place in that after a precisely specified measurement time, an insert which is provided with at least one surface, which can act as one or more boundary surfaces, to form one or more reaction openings, can be removed from the measurement arrangement.
• Platelets may have accumulated and aggregated there under the action of generated shear or flow forces on the boundary surface, which may or may not be bioactive or attachable.
• the characteristic curve is preferably formed such that it corresponds to a constant shear rate / force or flow rate.
• the size and type of the platelet reaction on the boundary surface can be optically evaluated after the platelet formation has been fixed, e.g. by means of an electron microscopic scanning system with subsequent computer-controlled image analysis and the display of measurement parameters for clinical diagnosis. Other optical evaluation options can also be used. (EP 0 635 720 A2).
• reaction device An embodiment is preferably used in which the adhesion and aggregate of the blood components (platelets) is caused under certain shear force conditions.
• a reaction opening can be used, as used in known devices for measuring the platelet function of primary hemostasis, for example in the form of an opening in a partition between the storage space and the pressure measuring device.
• Kunststoff EP 0 316 599 A2
• membrane EP 0 138 190 B1
• reaction openings are preferably used which are made entirely or partially of a hydrophilic material, for example polystyrene, glass or the like, or are surrounded by or consist of a bioactive material, in particular collagen (US Pat. No. 5,854,076 A; US 5,602,037 A; US 5,662,107 A).
• a very important aspect for the clinical acceptance of a procedure is, in addition to the delivery of clinically relevant data, its cost-effective application through inexpensive single-use measuring inserts and the possibility to carry out the measurements with small amounts of blood samples. Therefore, in the reaction devices of the corresponding design described, it is possible to use small amounts of blood for the measurement in such a way that, under the measurement conditions described, blood is transported from the storage space via the reaction device into the collection space and back again, resulting in global coagulation or platelet reactions , in particular the primary hemostasis in the reaction opening (s) until the parameter-forming measurement limits (time, volume, volume flow, pressure, formation formation for the optical evaluation etc.) of the respective measurement program have been reached, in order to then to be able to produce nosable results, as described above.
• the parameter-forming measurement limits time, volume, volume flow, pressure, formation formation for the optical evaluation etc.
• the blood viscosity in the starting phase can be advantageously determined via the precisely dimensioned geometry of the flow openings, the set volume flow and the resulting delivery pressure via the computer-controlled control mechanism.
• the correct shear rate can thus be set automatically and the influence of viscosity largely corrected as the measurement continues.
• Figure 1 shows an embodiment of a measuring arrangement for detecting coagulation functions of the global, in particular the primary hemostasis in whole blood or platelet-rich plasma.
• FIG. 2 shows an enlarged illustration of an embodiment of a reaction device for detecting coagulation functions of global, in particular primary, hemostasis, which can be used in the exemplary embodiment in FIG. 1;
• FIG. 3 shows a further embodiment for a reaction device for detecting coagulation functions of global, in particular primary, hemostasis, which can be used in FIG. 1;
• FIG. 4 shows a further embodiment for a reaction device which can be used in the device according to FIG. 1 and which corresponds to FIGS. 2 and 3;
• 4a and 4b show an embodiment for a reaction point which can be used in the reaction devices of FIGS. 3 and 4;
• FIG. 5 shows a further exemplary embodiment of a reaction device which can be used in the device according to FIG. 1;
• FIG. 6 shows a further exemplary embodiment of a reaction device which can be used in the arrangement according to FIG. 1;
• Figure 7 shows an embodiment for a reaction site
• Embossing in the vessel bottom / intermediate bottom and / or insert for forming flow openings which in the exemplary embodiments according to FIGS. 3 to 4; 5; 6; 17 and 19 can be used;
• Figure 8 shows an embodiment for a reaction site
• FIG. 11 shows a further exemplary embodiment of a reaction device for detecting coagulation functions of global, in particular primary, hemostasis, which can be used in the device according to FIG. 1;
• FIG. 12 shows a further exemplary embodiment of a reaction device which can be used in the device according to FIG. 1;
• reaction sites in particular reaction openings, which in the reaction device according to FIG. 12; 13; 16; 18 and 22 can be used;
• 13 to 24 show further exemplary embodiments of measuring arrangements for examining blood, in particular for detecting the platelet function of the primary hemostasis;
• FIG. 25 graphical representations of measurement results which are achieved with the exemplary embodiments of the measurement arrangements;
• FIG. 26 graphical representations of various possible vision / shear rate characteristics;
• FIG. 27 shows a graphical representation of a characteristic curve for regulating the volume flow as a function of
• FIG. 28 measurement results of hemostasis functions.
• FIG. 29 measurement results of hemostasis functions.
• a storage space 15 for blood to be examined is provided in a storage vessel 2.
• For the examination e.g. to capture the
• the reaction device 39 has a reaction point, e.g. Reaction opening 5 or a reaction channel, for which or which different embodiments can be provided. These embodiments are explained in more detail below.
• the respective reaction point (reaction opening) of the reaction device 39 can be designed such that blood components, in particular platelets, adhere there and aggregate and block the reaction opening partially or completely.
• the flow cross section provided in the reaction opening is narrowed and the flow resistance increases.
• a delivery pressure which corresponds to the pressure difference between a pressure generated by a delivery device, in particular suction pressure, and the pressure on the outside (atmospheric pressure) acts on the blood to be examined in the storage space 15.
• This delivery pressure changes at the narrowing of the cross-section of the reaction opening as a result of the possible accumulation and aggregation of the platelets or by a reduction in the flowability of the blood, by the onset of global, in particular primary, blood coagulation and an increasing flow resistance.
• a piston-cylinder arrangement 1 is used to generate the delivery pressure, which acts in a working space 12 on one side of the reaction device 39.
• This has a cylinder 25 (measuring cylinder) in which a piston 4 is axially displaceably guided.
• the motor 17 can be connected to the piston 4 via a coupling 13.
• the clutch 13 is releasable so that the piston-cylinder arrangement 1 can be separated from the engine 17.
• a pressure measuring space 3 is located below the storage space 15.
• the pressure measuring space 3 is located within a pressure-tight space into which the blood to be examined after passing through the
• Reaction device 39 arrives.
• this pressure-tight space is located in the working space 12 of the piston-cylinder arrangement 1.
• the blood that has passed through the reaction opening 5 and the central flow opening 11 of the reaction device 39 collects on a working surface 33, which in the exemplary embodiments shown in FIGS 17 is directed upwards.
• the piston 4 is sealed off from the inner wall of the cylinder 25, so that the amount of blood accumulating can be used as a measure of the volume flow rate.
• a liquid meniscus forming on the cylinder wall provides an additional gas-tight seal.
• the cylinder 25 can thus simultaneously serve as a measuring cylinder, since the movement of the piston 4 can be regulated by the drive 17 by the control unit 18 and thus the Volume flow and the volume flow rate through the reaction opening, can be detected as a measured variable.
• the pressure measuring chamber 3 is located above the surface of the blood in the cylinder 25, which has passed through the reaction device 39.
• the pressure measuring chamber 3 is free of the blood to be examined.
• a pressure line 8 designed as a hollow needle protrudes into the pressure measuring space 3. It is inserted gas-tight through the wall of the cylinder 25.
• the hollow needle 8 can have a tip that has pushed through the plastic material of the cylinder 25.
• a lifting magnet 19 can be used for this purpose, with which the hollow needle 8 can be pushed through the wall of the cylinder 25. This results in a pressure-tight passage of the hollow needle 8 through the cylinder wall.
• a pressure measuring device (pressure sensor) 9 is connected to the hollow needle or pressure line 8.
• the pressure line 8 can also be connected to the pressure chamber 8 in a pressure-tight manner in another way, for example via a connecting piece.
• the pressure measuring device 9 generates measurement signals which correspond to the pressure prevailing in the pressure measuring chamber 3.
• a pressure sensor can be arranged in the pressure measuring chamber 3, which generates corresponding measurement signals that can be transmitted wirelessly or with electrical connecting wires.
• the pressure measuring device 9 is connected to a control unit 18.
• the control unit 18 controls the drive of the motor 17 and thus the drive of the piston 4 as a function of the measured pressure prevailing in the pressure measurement chamber 3.
• the motor drive can optionally move the piston 4 in only one direction during the entire measurement process, namely from the upper position shown in Figure 1 to a lower position.
• the blood is pulsed back and forth through the reaction device 39 under predetermined shear conditions in the reaction or shear openings.
• the hollow needle 8 can also be connected to a vent valve 20 in order, if necessary, to be able to change the air cushion 62 in the pressure measuring chamber 3 via the movement of the piston 4.
• the piston 4 which limits the pressure-tight blood collection space 10 for collecting the blood which has passed through the reaction device 39 with its working surface 33, can be brought into a reference position before the start of the measurement by the Working surface 33 is moved against this stop before starting a measurement.
• the storage vessel 2 and the cylinder 25 are preferably made from one piece.
• the storage vessel 2 and the cylinder 25 can be of one
• Heating sleeve 23 are included, the temperature of which can be regulated.
• reaction openings 5 are designed in such a way that an accumulation of the blood components, in particular platelets, on the boundary surface 29 of the insert part 14 is initiated by the action of controlled shear forces.
• the insert 14 can be removed from the storage vessel 2 and the attached platelet formation 28 z. B. be examined optically, electron microscopy, chemically or physically to form clinical, diagnostic measurement parameters.
• the reaction device 39 has a reaction point or opening 5, which is formed by two opposing boundary surfaces 29 and 30.
• the boundary surface 29, which is preferably flat, is located on an insert 14 in the form of a stamp.
• the delimitation surface 30 is located on the vessel bottom 31 of the storage vessel 2.
• the two delimitation surfaces 29 and 30 can run parallel or not parallel to one another, the distance from the outside increasing slightly.
• the distance between the two boundary surfaces 29 and 30 defines the height of the reaction opening 5. In the exemplary embodiment shown, this extends essentially horizontally, ie perpendicular to the direction of movement of the piston 4.
• the delivery pressure generated by the piston-cylinder arrangement 1 located underneath it in the working space 12 acts via a central opening 11 and the reaction opening 5 on blood located in the storage space 15 of the storage vessel 2.
• the central opening 11 can also be formed by the interior of a tube 16 which can be inserted into the central flow opening 11.
• the central flow opening 11, or the tube 16, can be used as a feed opening and / or shear opening.
• the blood is transported from the outer edge of the reaction opening 5 to the central flow opening 11 or the tube 16.
• the boundary surface 30 has hydrophobic properties so that no platelet reaction can occur there.
• the stem-shaped insert 14 is arranged with the help of locking and spacer webs 24 in the correct position in the storage vessel.
• the slot height of the reaction opening 5 is determined by the spacer function.
• the boundary surface 29 is formed as a reaction surface.
• this can be coated or formed accordingly.
• the underside 32 of the insert 14, which forms the boundary surface 29, or the entire insert 14 can consist of hydrophilic material, for example polystyrene or glass; platelets can adhere to such materials.
• the underside 32 can also be kept hydrophobic and / or be provided with a roughness pattern.
• a bioactive coating 35 can also be provided on the underside surface 32 of the insert 14 in the form of an extracellular matrix (ECM), thrombin, batroxobin, collagen (also natural recombinant collagen or purified collagen).
• ECM extracellular matrix
• thrombin thrombin
• batroxobin collagen (also natural recombinant collagen or purified collagen).
• Subtypes synthetic peptides with collagen-like amino acid sequences, laminin, thrombospondin, fibronectin, blood cells, especially erythrocytes, leukocytes, preferably of the blood group zero, von Willebrand factor or from a mixture of collagen (as above) or synthetic peptides, each with substances such as Adenosine diphospate (ADP) or adrenaline or thrombospondin or fibronectin or other bioactive substances such as are listed, for example, in US Pat. No. 5,854,076A or US Pat. No. 5,662,107A, so as to form a bioactive boundary surface 29 of reaction opening 5. Furthermore, the boundary surface 29 can be formed in this way are, as shown in Fig.
• This occupancy can be coated or soaked with the bioactive substances specified above.
• the bioactive film or membrane can also consist of collagen.
• the coating can be designed differently. If necessary, several measurement results can then be obtained simultaneously. The different coverage / coating can be done in sectors or halves.
• 2a shows the boundary surface 29 with attached platelets after a measurement has been carried out.
• 2b shows the vessel bottom 31 as a top view for clarification.
• FIGS. 3 and 4 just like in FIG. 2, a change in the fluidity of the blood during blood coagulation (global hemostasis) can occur in the reaction opening (s), otherwise the platelet reaction of the primary hemostasis as in FIG. 2 should also be preferred on the boundary surface 29 of the insert 14, both of which (29 and 14) can be carried out according to the description in FIG. 2 (material and bioactive coating / coating).
• the insert 14 is also removable.
• the embodiment of FIG. 4 differs from FIG. 3 only in that the insert 14 with the surface 32, which forms the boundary surface 29, is designed as a flat cone and the vessel bottom 31, with the support surface 51, has been adapted to this shape ,
• impressions 53 are formed in a cross shape in the bottom of the vessel 31 (FIG. 4b). These embossments 53 form depressions between the support surfaces 51, on which the insert 14 is attached the surface 32, which forms the boundary surface (s) 29 of the reaction opening (s), lies in a form-fitting manner. This results in reaction openings 5 arranged in a cross shape, which essentially extend radially and horizontally (FIG. 3), and obliquely extending reaction openings 5 (exemplary embodiment in FIG. 4). On the boundary surface 29, which can be formed with the surface 32 of the insert 14, the cruciform deposits 28 of the platelets shown in FIG. 4a then result.
• the surfaces of the vessel bottom 31; Intermediate floor 50 and all surfaces which are formed by the impressions 53; 52 have arisen there, can optionally be made hydrophobic in order to avoid undesired platelet deposits there.
• FIGS. 5 and 6, 17 and 19 show further exemplary embodiments of reaction devices 39, in which flow openings are created by connecting an insert 14 and its surface 32 to the support surface 51 of the vessel base 31 or intermediate base 50 to its support surface 54 21 are generated, which reaction openings 5 or feed openings 27 and shear openings 46 can form.
• Embossments 53 are preferably formed in the vessel base 31 or intermediate base 50, as are shown, for example, in FIGS. 4b, 7 and 8. These impressions 53 form the bearing surface 51 onto which the insert 14 with its surface 32 is positively joined, this surface 32 representing a boundary surface 29 of the impression 53, as a result of which a flow opening 21 could be formed, as shown in FIG. 10a.
• the surface 32 of the insert 14 can also be designed such that it represents a boundary surface 29, as is described with FIGS. 2c to 2e, which creates a flow opening with the impression 53, which acts as a reaction opening 5, in which possibly cling and aggregate blood platelets.
• the flow opening 21 can also be formed such that an indentation 52 is only in the insert 14 and there creates a support surface 32 which is positively fitted onto the support surface 51 of the vessel base 31 or support surface 54 of the intermediate base 50 and so one Boundary surface 30 or 54, the indentation 52 is formed to form a flow opening 21, as can be seen from Figure 10b.
• the boundary surfaces of the flow opening 21 can then according to FIGS. 9a; 9b; 9c and their description.
• the impressions 52 can also be chosen arbitrarily in number and course and can also correspond to those in the intermediate base 31, according to FIGS. 4b; 7 and 8.
• impressions 52 in the insert 14 and impressions 53 in the vessel base 31 or intermediate base 50 can each be mirror images, which then form the mutual contact surfaces 32 and 51 or 54, which are positively joined onto one another in such a way that the impressions 52 and 53 form a flow opening 21 in a precisely covering manner, as shown by way of example in FIG. 10c.
• the boundary surfaces of the flow opening 21 can then according to FIGS. 9a; 9b; 9c and their description.
• the cross sections of the flow openings 21 of FIGS. 10a to 10c are shown by way of example and can also be shaped differently or interchanged.
• the number, type and shape of the flow openings 21, such as FIGS. 4b, 7, 8 and 9a to 9c, can also be selected differently according to the requirement.
• the flow openings 21, the design of which has been described for the reaction devices of FIGS. 5 and 6, 17 and 19, can perform different functions due to the surface texture / coating produced in each case.
• FIG. 9b in the flow opening 21, a predetermined area, the boundary surface 6, is provided with a bioactive coating 35, as a result of which a reaction opening 5 has been formed, while the upstream and downstream areas represent hydrophobic boundary surfaces 6, which thus supply openings 27 and can simultaneously form shear openings 46.
• FIG. 9c shows a flow opening 21, the boundary surface 6 of which has received a bioactive coating 35 over its entire length, and thus forms a reaction opening 5, at which plates can react with the aid of shear forces.
• the feed openings 27 and shear openings 46 can have a different opening cross section than the reaction opening 5 in the exemplary embodiments of the aforementioned figures.
• the bioactive coating 35 can consist of collagen (also natural recombinant collagen), synthetic peptides with collagen-like amino acid sequences, purified collagen subtypes, laminin, fibronectin, thrombospondin and other substances, or a mixture of collagen (as above) or synthetic peptides with each Adenosine diphosphate (ADP), adrenaline,
• Fibronectin, thrombospondin or other platelet activating substances (US 5,854,076A) (US 5,662,107A).
• the insert 14 and the vessel bottom 31 are predominantly made of Plastic, wherein the cross-sectional shape of the insert 14, which forms the surface 32 in addition to the shapes shown in FIGS. 2 to 4, can also be arcuate or tapering downwards, in which case the vessel bottom 31 with its boundary surface 30 or support surface 51 is adapted to these shapes.
• the insert 14, as seen from above, in addition to the round shape, can also have a triangular, oval, square and polygonal shape, and correspondingly also the receptacle in the vessel bottom 31.
• An insert receptacle can be formed on the vessel bottom 31 and / or on the vessel wall of the storage vessel 2 be in the form of locking and spacing webs 24 in which the insert 14 is inserted.
• FIG. 5 largely corresponds to the embodiment of FIG. 3, but it is not provided that the insert 14 can be removed from the storage vessel.
• the design options of the reaction device 39 correspond to FIGS. 4a and 4b, 7 and 8, 9a, b, c and 10a, b, c and their previous description.
• a tube 16 or hollow needle 55 can be inserted, which can be made of steel, plastic or glass and the flow surfaces of which are designed to be hydrophobic, so that shear effects (shear) can be generated there without causing platelet attachments can come.
• reaction openings 5 and 7, reaction device 39 platelet deposits occur under the influence of these precharging forces. This applies to all embodiments of FIGS. 2 to 24.
• the intermediate base 50 is formed on an extension of the vessel base 31.
• the intermediate floor 50 contains the central opening 11, via which the blood to be examined is introduced from the storage space 15 into the reaction device 39.
• the blood flows from the central opening 11 to the center outside towards the edge of the reaction device.
• the design options of the reaction device 39 also correspond to FIGS. 4a and 4b, 7 and 8, 9a, b, c and 10a, b, c and their previous description.
• a tube 16 can be inserted into the central opening 11 of the intermediate floor 50, in which the blood may be sheared according to the description of FIG. 5.
• FIG. 19 the same reaction device 39 is provided.
• the illustrated exemplary embodiments of the measuring arrangement can have the following dimensions in the order of magnitude.
• the diameter and the height of the storage vessel 2 can be approximately 10 to 20 mm.
• the height of the measuring cylinder 25 can be approximately 20 to 50 mm.
• the diameter of the measuring cylinder 25 can be approximately 8 to 15 mm.
• the flow opening 21 in the central flow opening 11 can have a diameter of approximately 0.300 to 3 mm, in the tube 16 approximately 0.100 to 2 mm.
• the length of the central flow opening 11 and the tube 16 can each be 0 to approximately 35 mm.
• the volume present in the pressure measuring chamber 3 can be approx. 10 to 50 ⁇ l, but in the FIGS. 18 to 24 approx. 500 to 1000 ⁇ l.
• FIG. 11 In the embodiment shown in FIG. 11 is located in a molded on the bottom 31 of the vessel
• Extension tube 16 which can be made of plastic, glass or steel.
• the flow opening 21 of the tube 16 can be designed as a reaction opening 5 and lined with a bioactive coating 35 as described in FIG. 9c. According to Figure 9 b and their
• the flow opening 21 can also be divided into a dimensioned section, which forms the reaction opening 5, and one or two remaining sections, which can form supply openings 27 or shear openings 46.
• the area that represents the reaction opening 5 may vary also located at the entrance in the middle or at the exit of tube 16.
• the tube preferably consists of polystyrene or glass, in which case the boundary surface 6 of the flow opening 21 is designed to be hydrophilic (in which case the bioactive coating can be omitted), and thus forms the reaction opening 5 according to FIG. 9a and its description.
• the diameter of the flow opening 21 of the tube 16 can be approximately 0.15 to 2 mm.
• the length can be about 10 to 30 mm.
• the tube 16 is removably arranged in the extension, so that an evaluation after the measurement is possible outside the measuring arrangement.
• the piston 4 is pot-shaped at its upper end and surrounds the downward-directed extension formed on the vessel bottom 31, in which the tube 16 is located, in order in this way to form the smallest possible air cushion 62 in the pressure measuring chamber 3 can.
• a reaction opening 7 is provided at the lower end of the tube 16 running in an extension of the bottom of the vessel.
• the flow opening 21 of the tube 16 can act as a feed opening 27 and / or a shear opening 46.
• the reaction opening 7 can be designed as is known from EP 0 111 942 or EP 0 316 599 A1.
• the reaction opening 7 or partition wall 34 can preferably be designed as shown in FIGS. 12a to 12d.
• the reaction opening 7 is located in a partition 34 made of plastic, for example polystyrene, with hydrophilic surfaces or made of bioactive material, which can be attached to platelets, for example in the form of a bioactive film or a non-porous collagen membrane.
• the partition wall 34 consists of non-porous material which is attached to one of the two partition walls or to both partition surfaces and / or to the boundary tion surface 6 of the reaction opening 7 is provided with a bioactive coating 35, as described below.
• both partition walls of the partition 34 and the boundary surface of the opening are provided with a bioactive coating 35.
• the bioactive coating 35 / impregnation can consist of collagen (also natural recombinant collagen or purified collagen subtypes), synthetic peptides with collagen-like amino acid sequences, laminin, fibronectin, thrombospondin or other substances, bioactive substances (US Pat. No.
• the reaction opening 7 is located at the lower end of the tube 16, which can have a length of 0 to approximately 35 mm and a diameter of approximately 0.150 to 2 mm.
• the partition wall 34 with the reaction opening 7 can also be provided approximately in the middle of the tube 16 in the direction of its longitudinal axis (not shown).
• the partition wall 34 with the reaction opening 7 can also be arranged in a recess in the vessel bottom 31 without the tube 16.
• the opening diameter of the reaction opening 7 can be about 0.100 to 0.500 mm.
• the wall thickness of the partition can be about 0.10 to 6 mm.
• the storage space is located in the cylinder 59, preferably in a blood collection syringe.
• the reaction device 39 is essentially designed as it is shown in FIG. 12, but it can also be designed as it is shown and described in FIGS. 6 and 11.
• the blood to be examined in the storage space 15 of the syringe barrel 59 of the withdrawal syringe 45 flows through a hollow needle 55, which is fastened in a pressure-tight manner in the housing 64 and with its free needle end 47 projecting through it, is inserted through a sealing insert in the syringe adapter 22 of the syringe barrel 59 Reaction device 39 in the blood collection space 10 located below.
• the piston 4 is moved downward along the measuring cylinder 25 to carry out the measurement.
• the piston 4 in the measuring cylinder 25 can optionally move the blood back and forth in the reaction device 39 under measuring conditions.
• the hollow needle 55 is completely hydrophobic on the surfaces and can consist of steel, plastic or glass, and its flow opening can act as a feed opening 27 and a shear opening 46. This embodiment of the hollow needle 55 can also be used identically in FIGS. 14 and 15, 22 to 24.
• the reaction opening 7 is preferably designed as described with reference to FIGS. 12a and 12b.
• the pressure measurement in the pressure measuring chamber 3 takes place via the pressure line 8, as has already been explained in the previously described exemplary embodiments.
• the hollow needle 55 is fixedly connected to a housing 64 in which the reaction device 39 is accommodated. Aeration is required for an air cushion 62 above the blood sample located in the storage space 15. device / vent 61 provided so that no back pressures can build up in this space during the measuring movement of the piston 4 in the piston-cylinder arrangement. 1
• the hollow needle 55 is also attached to the housing 64 of the reaction device 39.
• the reaction device 39 can also be designed in the same way as in the exemplary embodiments of FIGS. 6, 11 to 13.
• the hollow needle 55 is connected via a sealing sleeve 56 with a catheter 57, for the direct measurement of the patient's blood e.g. from the vein.
• the storage space 15 for the blood to be examined is in a container 65, which can be placed on the hollow needle 55 attached to the housing 64, the tip of the free needle end 47 in in the bottom region of the container 65 has hit the storage room 15.
• the penetrated material in the bottom area of the container 65 lies in a sealing manner against the hollow needle 55, so that the desired transport of the blood to be examined through the reaction device 39 takes place during the movement of the piston 4.
• this embodiment is suitable for measuring pipetted blood, which e.g. Substances have been added or which has been manipulated in some other way or if only a small amount of blood is available for a control measurement. (E.g. below 500 ⁇ l).
• the reaction device 39 can also be designed as shown in FIGS. 6, 11 to 14.
• the piston / cylinder arrangement of a special syringe (not shown) or a withdrawal syringe (disposable part) serves as a conveying device in order to draw the blood to be examined out of the working space 12 of the conveying device, which Room 15 is used to transport the hollow needle 55, which is fixed pressure-tight in the central flow opening 11, and then through the reaction device 39.
• the vessel located at the top comprises the blood collection space 10.
• the piston 4 present in the syringe cylinder 59 is connected to the drive 17 via a coupling (not shown in more detail).
• the piston 4 in the syringe cylinder 59 can be moved upward in one direction by the electrical drive device 17, in accordance with the measurement program regulated by the control unit 18, in order to convey the blood through the reaction device 39.
• the piston 4 can also be moved back and forth so that the blood flows from changing directions through the reaction opening 7, which is located in the partition 34, which can be designed according to FIGS. 12a and 12b and the description in FIG.
• the partition wall 34 can be located in a recess in the vessel bottom 31 of the vessel 2 in question.
• reaction devices 39 can also be used, which correspond to the embodiments of FIGS. 2 to 5, 11 and 17 and the explanations therefor.
• Anticoagulated blood drawn from the patient is filled with the reaction device 39 in order to carry out the measurement process, the free needle end 47 of the hollow needle 55, which projects downward, is guided in a syringe adapter through the sealing insert 26 in a pressure-tight manner.
• the interior of the syringe adapter 22 of the withdrawal syringe 45 forms the pressure chamber 3.
• the other end of the hollow needle 55 is fixedly connected to the vessel bottom 31 in the central flow opening 11.
• the removal syringe 45 forms the piston-cylinder arrangement 1, the piston 4 of which has a Coupling 13, not shown, is connected to the drive 17.
• the piston 4 is moved upwards in the cylinder 25, in the direction of the reaction device 39 and, with its working surface 33, pushes the blood column formed by the inner wall of the cylinder 25 upwards when the vent valve 20 is open, until the upper blood level is at a precisely defined one Position is detected by the level sensor 58, which then signals the control unit 18 the reference position, from which the measurement can then start.
• the reference position defines the volume of the air cushion 62, which forms the blood-free pressure measurement space 3 and is limited by the outer surface of the hollow needle 55, the blood level, the inner wall of the syringe adapter 22 and the underside of the sealing insert 26.
• FIG. 17 shows an embodiment with a reaction part 39, which can correspond to that of FIGS. 2 to 5. Otherwise, the embodiment is identical to that described in FIG. 16.
• the delivery device for generating the delivery pressure outside the common housing 38.
• the delivery device consists of a piston / cylinder arrangement with the cylinder 40 and the piston 4.
• the piston 4 is like in the exemplary embodiments described above driven by the electric drive device, for example a stepper motor 17.
• the pressure line 8 which can be designed as a hollow needle and the tip of which has pushed through the container wall 37 of the blood collection space 10 by means of a drive 19, as described in FIG. 1, carries the delivery pressure generated in the working space 12 of the piston / cylinder arrangement. This pressure corresponds to the pressure to be measured in the pressure measuring room 3.
• a pressure measuring line 41 is branched off from the pressure line 8 and connected to the pressure measuring line 9 connected.
• the pressure measuring device 9 (sensor) is connected to the control unit 18, to which the electric drive device 17 is also connected.
• the blood collection space 10 is located in the common housing 38 below the storage vessel 2.
• the reaction device 39 can be designed as shown in the exemplary embodiments in FIGS. 12 to 15. A further possibility is that the reaction device 39 can, preferably, be designed as a separate insert and can be inserted into the common housing 38 from the storage vessel 2 and the blood collection vessel 66 in such a way that it lies on the limiting webs 44.
• the outer ring of the part of the reaction device 39 forming the bottom 31 of the vessel is designed such that it can act as a sealing element 66 to the inner wall of the reservoir 2.
• This embodiment can optionally also be used in other embodiments.
• a blood sensor 67 is located below the reaction device 39 in the blood collection space 10. This sensor detects the first drop of blood that has passed through the reaction device 39 and a converter 63 then sends a signal to the control device 18 that the measurement is ready to start.
• a reaction device 39 is used, as is used in the exemplary embodiment in FIG. 6.
• a reaction device is used, as described in the embodiment of FIGS. 3 and 5.
• the storage space 15 for the blood to be examined is located in the cylinder of a withdrawal syringe 45.
• the reaction device 39 is located in the housing 64, which, as in FIG 18 shows, in its lower part, the pressure-tight blood collection space 10.
• the piston-cylinder arrangement 36 by means of which the measuring pressure is built up, serves to convey the blood through the reaction device 39 can be designed as shown in FIG. 18.
• a hollow needle 43 which is connected to the withdrawal syringe 45 via the cannula adapter 60, can be pushed through a wall region of the housing 64, as shown in FIG. 21. This creates a pressure-tight connection with the reaction device 39 inside the housing.
• the measurement process during the examination of the blood and the combination with reaction devices 39 of other figures and the execution of parts then takes place in the manner as was explained in the exemplary embodiments described above, in particular with FIG. 13.
• For an air cushion 62 above that in FIG A blood supply / vent 61 is provided in the storage chamber 15 so that no back pressures can build up in this space during the measuring movement of the piston 4 in the piston-cylinder arrangement 36.
• a piston-cylinder arrangement 1 integrated in the housing 64 can also be used, as is the case in the exemplary embodiments in FIGS. 1 to 17 is used. It is also possible to use the possible reaction devices 39 described and named for FIGS. 13 and 22.
• the exemplary embodiment shown in FIG. 22 is used using the external conveying device in the form of the piston-cylinder arrangement 36 in accordance with FIG. 18, and the blood collection space 10 is also located in a container closed at the bottom, as in FIG. Otherwise, this embodiment is largely identical to Figure 13 and its description.
• the measuring arrangement shown in FIG. 23 largely corresponds to that in FIG. 14 and the measuring arrangement shown in FIG. 24 largely corresponds to the measuring arrangement shown in FIG. 15, but can be used in each case according to FIG. 18, as described in FIG. 22
• the embodiment in FIG. 24 is also suitable for measuring pipetted blood, in contrast to FIG Substances have been added or which has otherwise been manipulated or if only a small amount of blood is available for a control measurement (e.g. below 500 ⁇ l).
• the detection of the platelet function of the primary hemostasis can also be carried out in such a way that the measured pressure is maintained by feedback to a setpoint pressure value and as a measure of the aggregation or coagulation of the platelets by the amount of blood flow the capillary is determined (DE 35 41 057 AI).
• the detection method in such a way that the pressure change which forms in the reaction device 39 during the progressive addition of the respective flow path is measured at certain time intervals and the volume flow is changed in each case that it corresponds to a predetermined function. Also during pre- At certain intervals, the pressure is kept constant and then, when the volume flow has dropped by an amount, readjusted until it corresponds to the specified function (DE 196 17 407 AI).
• a new method according to the invention is preferably used, in which, depending on the pressure measured in the pressure measuring space 3, the volume flow of the blood to be examined is adjusted by the reaction device 39 such that a predetermined shear rate or shear force characteristic is maintained and preferably the shear rate or shear force is kept constant.
• the volume flow rate and / or the then available volume flow after a given measuring time or the given time or at a given volume flow rate, the elapsed time and / or the then existing volume flow or with a given volume flow, the elapsed time and / or then existing volume flow rate can be used.
• the pressure increase after a predetermined time or the elapsed time after the specification of a pressure increase can be used for parameter formation.
• volume flow is set according to the following relationship:
• V the volume flow of the blood to be examined through the reaction device, in particular through the shear opening;
• the measurement arrangement in particular the piston movement, can be controlled in such a way that a blood flow runs along a predetermined shear rate or shear force characteristic.
• FIG. 26 shows a non-linear, increasing shear force characteristic and a decreasing linear characteristic in dash-dotted form.
• the course of the respective characteristic curve for the shear rate (1 / s) or the shear force (N / m 2 ) can optionally be selected as a function of the diagnosis to be made, for which the measurement is carried out.
• a constant characteristic curve (solid line in FIG. 26) is selected for a specific shear rate or shear force. If the flow pressure increases, for example due to the accumulation or aggregation of the platelets in the reaction opening, the desired characteristic curve is achieved by controlling the piston movement in accordance with the formula given above.
• FIG. 27 shows the volume flow normalized to 1 at 5 mbar as a function of the pressure difference dp for regulating the volume flow as a function of dp in the reaction opening when a constant shear rate is specified.
• 28 and 29 show measurement results including the pressure curve when checking the coagulation function of the global, in particular the primary hemostasis when regulating the volume flow as a function of the pressure change in the reaction opening, with a constant shear rate according to FIG. 27 Evaluation, are the shutter speed and the shutter volume. 28 the volume flow goes to zero in 196 seconds and a volume flow rate of 310.9 ⁇ l.

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• 2000
  • 2000-01-25 DE DE10003093A patent/DE10003093C2/de not_active Expired - Fee Related
• 2001
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