DE102016106097B3 - Tissue and organ transport device - Google Patents

Abstract

Tissue and organ transport device (1) for the transmission, transport and stabilization of tissues and organs (6), comprising a solid container (2) with a cover (7) and a lower vessel (3 '), wherein for the tissue ( 6) or the organ (6) a macroporous support material (4) is embedded with a system of interconnected pores having a pore size of ≥ 1 μm fixed in the lower vessel (3 ') and allows a hydrodynamic flow and the lower vessel (3') is formed with a movable piston (5) for generating a hydrodynamic pressure.

Description

The invention relates to a tissue and organ transport device for the transmission, transport and stabilization of tissues and organs. The device is intended for use for stabilization, gentle transport, and transmission of tissues and organs from the preparation site to the patient to receive the tissue or organ. In addition, the tissue and organ transport device is applicable for accurate and gentle positioning of tissues and organs during surgery, particularly transplantation or implantation.

A prerequisite for successful transport of tissues and organs is to maintain a certain temperature, depending on the tissue or organ that is to be transported. Maintaining the temperature is important to ensure that the tissue or organ is not heated or frozen too early and is damaged.

Organs, including the heart, lungs, liver, pancreas, and kidney, must be transplanted immediately after explantation, as currently no suitable organ culture techniques are available. Therefore, the organs should be rapidly cooled to 4 ° C and kept in this temperature range for periods up to 24 hours. They are placed in special foil pouches and stored in a styrofoam container filled with crushed ice.

Tissues, such as the amniotic membrane (egg skin), heart valves and vascular valves, are cryopreserved immediately after collection and must be placed on dry ice. In contrast, donated corneas are cultured at 31 ° C to 37 ° C in growth medium. For transport from the tissue bank to the surgeon, the corneas are placed in simple chambers with a fastening system that protects the tissue from swimming around during transport, for example, into a so-called Böhnke chamber with a corneal retainer.

In addition, for example, there is no transport system at all for precut endothelial cells of donor cornea because they are very thin, that is, about 100 μm, and the fragile tissue remains in the bed of the donor cornea to transport it from the tissue bank to the surgeon.

From the JP 2002-335 950 A a container for storing and transporting a membrane tissue is described, which encloses a biological material. In this case, the container comprises a liquid-permeable support structure based on, for example, a sponge-like material which encloses the flat membrane tissue.

The WO 02/026 034 A2 describes a device which maintains the ex vivo viability of organs and can be used for the storage and transport of organs. The device contains a variety of membranes, valves and piping. Furthermore, a manual or computerized control to maintain the system is necessary.

In the JP 2010-220 488 A An instrument is described that can be used to record cell culture products, such as cultured tissues. The instrument consists of a plastic container with a porous wall which, when contacted with the cell culture products, can pick them up for example by means of a vacuum and release them later by applying an applied pressure. This instrument is intended to facilitate the handling of the cells and their products in laboratories, but on the other hand does not allow storage or transportation outside of a sterile environment.

The DE 10 2005 004 826 A1 describes a suction cup for temporary removal of at least a portion of a cornea. The Saugtrepan includes, inter alia, a fluid-permeable contact body. This has an open-pored microporous material with a pore size of 0.6 to 1.2 microns. As possible materials for the plant body various ceramic materials are specified.

From the JP 2014-132 899 A a device is known which is suitable for receiving cell culture products. The device comprises a syringe cap with a porous wall made of plastic and / or glass, which is in contact with cell culture products and which can absorb these cell culture products by means of vacuum, transfer and release by injection. The device is intended to simplify the handling of the cells and their products under laboratory conditions. However, this device is not suitable for transporting biological samples outside of laboratories.

The object underlying the invention is to provide an adapted tissue and organ transport device which is suitable as a mechanical support for very thin and delicate tissue, including endothelial cells of the cornea, as well as for organs. This device must ensure the alignment of the tissue and must not result in or cause damage to the tissue or organ during transport. Such a transport device for tissue and Organs must ensure continued supply of nutrients and should also allow continuous supply of growth-promoting and viability-ensuring components.

In addition, the tissue and organ transport device should also allow maintenance of the temperature required during transport of the tissue or organ. The tissue and organ transport device should also allow the gentle delivery and positioning of implants during surgery, without having to touch the tissue or organ with tweezers or other surgical instruments.

The object of the invention is achieved by a tissue and organ transport device having the features of claim 1. Advantageous developments are specified in the subclaims.

The tissue and organ transport device is suitable for the transfer, transport and stabilization of tissues and organs, and comprises a solid container having a lid and a bottom vessel, wherein the tissue or organ is a macroporous carrier material having a system of interconnected pores embedded in the lower vessel and allows a hydrodynamic flow and the lower vessel is formed with a movable piston for generating a hydrodynamic pressure.

The invention is based on macroporous support materials having a minimum pore size of ≥ 1 μm, preferably of several, ie ≥ 2 μm, as well as a crosslinked pore system. The macroporous support materials are embedded in a solid container, for example by the application of surface functionalization, a specific surface topography, adhesives, seams or a special haptic geometry of the container. According to an advantageous embodiment of the invention, the macroporous support material is functionalized with chemical and / or biological entities.

The geometry of the tissue and organ transport device may be adapted to the tissue or organ that is to be transported, transplanted or implanted. For example, the tissue and organ transport device may be in the form of a flat, round cylinder, a tall round cone, or the shape of a syringe. By exerting hydrodynamic pressure, for example via a syringe system, the tissue or organ can be carefully "pushed" out of the device and applied to the site of implantation in the patient's body without much mechanical effort by the use of forceps and other surgical equipment.

In this context, the geometry of the macroporous support material may also be adapted to the shape and geometry of the tissue or organ that is to be stabilized, transported and transplanted / implanted. For example, the macroporous support material may have a cylindrical hole, a concave cavity or a convex shape. The macroporous support materials can be subjected to swelling via contact with aqueous media.

The macroporous support materials can be made by any type of material, including natural sponges, foams, pure synthetic or biological polymers, synthetic or biological polymer blends, ceramics, rubber, and other materials.

Among these materials, biohybrid hydrogels, as described, for example, in PB Welzel et al., 2012 Macroporous starPEG-heparin Cryogels, Biomacromolecules 13, 2349-2358, are preferred because they can carry (bi) functional units that enhance survival and survival Support growth of tissues and organs during transport and later implantation. In addition, the macroporous support materials can be prepared by known methods, including porogen leaching, described inter alia in W. L. Murphy et al. 2002, Salt fusion: an approach to improve pore interconnectivity within tissue engineering scaffolds. Tissue Eng. 8 43-52, the gas-foaming technique, as known, for example, from A. Sannino et al. 2006, Synthesis and characterization of macroporous poly (ethylene glycol) -based hydrogels for tissue engineering application. J. Biomed. Mater. Res. A 79 229-236, the phase separation described, for example, in T.D. Dziubla 2002 Macroporous Hydrogels as Vascular- Arisable Soft Tissue - Impant Interfaces: Materials Characterization, electrospinning, known inter alia from Q. Pham et al. 2006 Electrospinning of polymeric nanofibers for tissue engineering applications: a review. Tissue Eng. 12, 1197-1211, and cryogelation in aqueous media, described, inter alia, in I. Kaetsu 1993 Radiation Synthesis of Polymeric Materials for Biomedical and Biochemical Applications Adv. Polym. Sci. 105, 81-97. The invention provides a powerful transport tool that ensures the mechanical support of very thin and delicate tissue, including corneal endothelia, and organs. This mechanical hold is based on the cushioning effect of the macroporous support materials embedded within the solid container.

The tissue and organ transport device according to the invention ensures the maintenance of the order or orientation of the tissue and does not cause any damage to the tissue or organ during transport.

In addition, the tissue and organ transport device ensures sustained nutrient delivery and provides a continuous supply of growth-promoting and viability-ensuring components. In addition, the tissue and organ transport device ensures maintenance of the temperature required during transport of the tissue or organ.

Further details, features and advantages of embodiments of the invention will become apparent from the following description of exemplary embodiments with reference to the accompanying drawings. Show it:

1A : a schematic representation of a tissue and organ transport device according to a first variant,

1B : a schematic representation of a tissue and organ transport device according to a second variant,

1C FIG. 2 is a schematic representation of a tissue and organ transport device having regions for generating a hydrodynamic pressure. FIG.

1D FIG. 2 is a schematic representation of an alternative embodiment of the tissue and organ transport device according to the invention, which is in the form of a syringe, FIG.

2 An embodiment of an application of a macroporous support material based designed device for the transmission and transport of implantable tissues and organs, and

3 : A schematic representation of the transmission and transport of a sensitive tissue using a syringe-shaped embodiment of the tissue and organ transport device according to the invention.

The 1A shows a first variant of a tissue and organ transport device 1 which contains essential but not all features of the invention, this device 1 a solid container 2 with a lower vessel 3 in the form of a flat cylinder into which a macroporous support material 4 is embedded. On the surface of the macroporous support material 4 in the middle of the cylindrical lower vessel 3 is a cavity 5 for a tissue 6 or organ 6 educated. tissue 6 or organs 6 can thus be in or on the macroporous support material 4 put in a tight container 2 is embedded. Furthermore, the fixed container 2 according to 1A a lid 7 also in the form of a flat cylinder, in which no macroporous carrier material is embedded. The advantage of the macroporous support material is a cushioning effect on the tissue or organ. Furthermore, the porous carrier material allows 4 a light, gentle hydrodynamic flow. These two advantageous features are indicated by the double arrow in the detail section of the 1A indicated.

The 1B shows a second variant of a tissue and organ transport device 1 which contains essential but not all features of the invention, wherein the device 1 also a solid container 2 with a lower vessel 3 in the form of a flat cylinder into which a macroporous support material 4 is embedded. On the surface of the macroporous support material 4 in the middle of the cylindrical lower vessel 3 is a cavity 5 for a tissue / organ 6 educated. Tissues / organs 6 can thus be in or on the macroporous support material 4 put in a tight container 2 is embedded. In contrast to 1A is in the variant according to 1B also in the bottom of the lid 7 , which is formed in the form of a flat cylinder, porous support material 4 with a cavity placed in the middle 5 for the tissue / organ 6 embedded. As shown in 1B is this cavity 5 of the macroporous support material 4 in the lid 7 the cavity 5 of the carrier material 4 in the lower vessel 3 exactly opposite. After closing the solid container 2 envelop the cavity 5 in the macroporous carrier material 4 of the lower vessel 3 and the cavity 5 in the macroporous carrier material 4 of the lid 7 as a united cavity together the tissue / organ 6 , The advantage of the macroporous carrier material 4 consists in a cushioning effect on the tissue or organ. Furthermore, the porous carrier material allows 4 a light, gentle hydrodynamic flow. The advantages of using the macroporous substrate 4 , namely the padding effect and the gentle hydrodynamic flow, are in the detail of the 1B indicated schematically by the double arrow.

The 1C shows a third variant of a tissue and organ transport device 1 containing essential but not all features of the invention. In this case, the tissue and organ transport device comprises 1 a solid container 2 , which is a cylindrical lower vessel 3 and a lid 7 having. In the bottom of the lid 7 in shape a flat cylinder is formed, macroporous carrier material 4 with a cavity placed in the middle of the cylinder 5 embedded for the tissue / organ. The lower vessel 3 is not formed here in the form of a flat cylinder, but as a cylinder, which is below the carrier material 4 and the cavity 5 areas 8th for generating a hydrodynamic pressure, wherein in 1C the application of a hydrodynamic pressure in different directions by double arrows is shown schematically.

Finally, the shows 1D a fourth variant of the tissue and organ transport device according to the invention 1 , Wherein also according to this variant in the bottom of the lid 7 , which is formed in the form of a flat cylinder, macroporous carrier material 4 with a cavity placed in the middle of the cylinder 5 for the tissue / organ 6 is embedded. The lower vessel 3 ' is in the form of a syringe 3 ' with a movable piston 5 formed, via an operating element 10 in the cylindrical lower vessel 3 ' is movable in the axial direction. By an axial movement of the piston can along or parallel to the axial direction in the areas 8th that are in the lower vessel 3 ' below the carrier material 4 and the centrally placed cavity 5 with the tissue / organ 6 are located, a hydrodynamic pressure are generated.

• a) Makroporöse Materialien haben eine geringe Volumensteifigkeit und eine hohe mechanische Stabilität beim Zusammendrücken. Diese Eigenschaften ermöglichen eine einfache Handhabung der Materialien und ermöglichen eine schonende geometrische Anpassung an die im Allgemeinen unregelmäßige Form von Geweben und Organen. Diese hier als „Polsterungseffekt" bezeichnete Eigenschaft ermöglicht die schonende Stabilisierung, Behandlung, den Transport und die Abgabe von Geweben und Organen, ohne dass eine mechanische Kraft ausgeübt wird, die die Unversehrtheit des Gewebes und der Organe beeinträchtigen könnte.
• b) Bei Ausübung eines hydrodynamischen Drucks, beispielsweise über ein Spritzensystem, wie in 1D gezeigt, kann das Gewebe oder Organ ohne eine große mechanische Einwirkung, das heißt unter Verzicht auf die Anwendung von Pinzetten und anderen chirurgischen Geräten, vorsichtig aus der Vorrichtung 1 „geschoben“ und zum Beispiel an eine Implantationsstelle im Körper des Patienten angelegt werden.
• c) Makroporöse Materialien können mit jeder Art von Puffer oder Medium, einschließlich Organdurchblutungsmittel, Gewebe- oder Organkultur- beziehungsweise Wachstumsmedium gefüllt werden. Aufgrund des Systems von miteinander verbundenen Poren ist die kontinuierliche und ungehinderte Versorgung des Gewebe- oder Organs sichergestellt. In diesem Zusammenhang besteht ein wesentlicher Vorteil darin, dass das makroporöse Trägermaterial eine gleichmäßige Temperaturverteilung und eine gute Wärmeleitung gewährleistet.
• d) Darüber hinaus kann in Abhängigkeit von der Chemie des makroporösen Trägermaterials dieses mit chemischen oder biologischen Einheiten funktionalisiert werden, um das Überleben, die Überlebensfähigkeit und auch das Wachstum der zu transportierenden und implantierenden Gewebe und Organe zu unterstützen.

The macroporous structure of the carrier material 4 Having a system of interconnected pores has several advantages:

• a) Macroporous materials have low volume stiffness and high mechanical stability upon compression. These properties allow easy handling of the materials and allow a gentle geometric adaptation to the generally irregular shape of tissues and organs. This property, referred to herein as the "cushioning effect," allows the gentle stabilization, treatment, transport, and delivery of tissues and organs without exerting a mechanical force that could compromise the integrity of the tissue and organs.
• b) When a hydrodynamic pressure is exerted, for example via a syringe system, as in 1D The tissue or organ may be carefully removed from the device without a major mechanical impact, that is, without the use of tweezers and other surgical devices 1 "Pushed" and placed for example at an implantation site in the body of the patient.
• c) Macroporous materials can be filled with any type of buffer or medium, including organ perfusion agents, tissue or organoculture or growth media. Due to the system of interconnected pores, the continuous and unobstructed supply of the tissue or organ is ensured. In this context, a significant advantage is that the macroporous support material ensures a uniform temperature distribution and good heat conduction.
• d) In addition, depending on the chemistry of the macroporous support material, it may be functionalized with chemical or biological moieties to aid in the survival, viability and growth of tissues and organs to be transported and implanted.

The applicability of the tissue and organ transport device has been investigated for a very demanding task, namely the stabilization and transfer of a tissue engineered cellular monolayer, as in US Pat 2 shown schematically, wherein the 2 shows essential but not all features of the invention. This is the cellular monolayer 11 on a thermoresponsive cell culture carrier 12 that is, a carrier with a thermoresponsive polymer. Stimulable cell culture carriers 12 , for example, thermo-responsive cell culture carriers 12 , are for the culture of cellular monolayers 11 suitable for endothelial cells of the human cornea. By using an external trigger, for example a temperature reduction, a detachment takes place 13 the cellular monolayer 11 from the stimulable cell culture carrier 12 , This so-called cellular monolayer 11 is extremely thin, namely about 10 microns, and brittle and therefore can not be touched by pliers or raised. It is known that cell turfs that consist of only one cell layer can be stabilized using membranes including poly (vinylidene fluoride) and cellulose acetate membranes and transferred to a new substrate. The disadvantage is that these membranes are very stiff and are hardly applicable to substrates with a non-planar geometry, for example the concave curvature of the cornea.

With the help of macroporous biohybrid hydrogels 14 , so-called cryogels, could replace detached endothelial cell layers 11 stabilized on the human cornea and on swine corneas as target substrates 15 be transmitted. The cryogel structuring has been adapted to produce these sponge-like cryogels based on the chemical crosslinking of four-armed poly (ethylene glycol) (starPEG) and heparin.

For this purpose, an aqueous gel-forming reaction mixture composed of starPEG and heparin is frozen at temperatures below 0 ° C. (-20 ° C.). Under these conditions, the macroscopically frozen sample formed two phases, a polycrystalline phase of frozen pure water and a non-frozen liquid microphase containing high concentration starPEG and activated heparin. The crosslinking reaction proceeded in the non-frozen liquid microphase. Ice crystals acted as porogens, that is, as pore formers. They were removed from the frozen gels by lyophilization (sublimation), resulting in the generation of a highly porous scaffold structure.

Macroporous samples of various sizes or shapes were prepared by this method. For stabilization and transfer of cell layers, cryogel discs with a diameter of 1 cm and a thickness of 2 mm, based on the swollen state, were used. Cell detachment and cell transfer using cryogels did not affect the survivability of the human human corneal endothelial cell layers transferred. Cryogels showed a porosity of up to 92% with interconnected pores, low volume stiffness, and high mechanical stability upon compression. These properties allowed easy handling of the cryogels and a lifting of the detached cell layers by the "sponge effect" of this macroporous material. The cryogels allowed a good adaptation of the cryogels together with the transferred cell layer to the concave curvature of the pig's cornea. In addition, the high porosity ensures the continuous supply of nutrients to the transferred tissue.

In this connection, it should be noted that the cryogels can remain on the transferred cell layer for at least one day, which supports the stable attachment of the transferred cells to the new target substrate.

The 3 schematically shows the transmission and transport of the endothelial lamella as an example of a delicate tissue using a syringe-like tissue and organ transport device according to 1D , In method step I), a gentle application of the endothelial lamellae to the macroporous carrier material (gel) is initially carried out by generating a liquid flow by moving the piston in the suction direction. The macroporous gel is relaxed.

The transport takes place in method step II) while maintaining the orientation of the fabric due to the existing hydrodynamic pressure with the pistons drawn out.

Finally, in method step III), a precise, gentle delivery of the tissue is achieved by generating a liquid flow by moving the piston in the ejection direction. The macroporous gel is compressed.

LIST OF REFERENCE NUMBERS

1

 Tissue and Organ Transport Device, Device

2

 solid container

3

 lower vessel

3 '

 Lower jar in syringe form, syringe

4

 macroporous carrier material

5

Cavity in the carrier material 4

6

 Organ, tissue

7

 cover

8th

 Areas for generating a hydrodynamic pressure

9

 piston

10

Control element (for the piston 9 )

11

 cellular monolayer, endothelial cell layer

12

 stimulable cell culture carrier

13

Replacement of the cellular monolayer 11 from the stimulable cell culture carrier

14

 Biohybrid hydrogel

15

 target substrate

Claims (8)

Tissue and organ transport device ( 1 ) for the transmission, transport and stabilization of tissues and organs ( 6 ) comprising a solid container ( 2 ) with a lid ( 7 ) and a lower vessel ( 3 ' ), where for the tissue ( 6 ) or the organ ( 6 ) a macroporous carrier material ( 4 ) with a system of interconnected pores having a pore size of ≥ 1 μm in the lower vessel ( 3 ' embedded) and allows a hydrodynamic flow and the lower vessel ( 3 ' ) with a movable piston ( 5 ) is designed to generate a hydrodynamic pressure. Tissue and organ transport device ( 1 ) according to claim 1, characterized in that the macroporous support material ( 4 ) has a pore size of ≥ 2 microns. Tissue and organ transport device ( 1 ) according to claim 1 or 2, characterized in that the geometry of the tissue and organ transport device ( 1 ) to the tissue to be transported ( 6 ) or organ ( 6 ) is adjusted. Tissue and organ transport device ( 1 ) according to one of claims 1 to 3, characterized in that the geometry of the macroporous support material ( 4 ) to the shape and geometry of the tissue to be transported ( 6 ) or organ ( 6 ) is adjusted. Tissue and organ transport device ( 1 ) according to one of claims 1 to 4, characterized in that macroporous support material ( 4 ) is functionalized with chemical and / or biological moieties. Tissue and organ transport device ( 1 ) according to one of claims 1 to 5, characterized in that the macroporous support material ( 4 ) is swellable by contact with an aqueous medium. Tissue and organ transport device ( 1 ) according to any one of claims 1 to 6, characterized in that the macroporous support material ( 4 ) is prepared by a process selected from porogen leaching, solid-state foam formation, gas-foaming, phase separation, electrospinning and cryogel formation in an aqueous medium. Use of a tissue and organ transport device ( 1 ) according to one of claims 1 to 7 for the transmission, transport and stabilization of tissues or organs to be transplanted or implanted ( 6 ).

Images