WO2017174072A1

WO2017174072A1 - Tissue- and organ transport device

Info

Publication number: WO2017174072A1
Application number: PCT/DE2017/100261
Authority: WO
Inventors: Mikhail Tsurkan; Juliane Teichmann; Carsten Werner; Petra Welzel; Mirko Nitschke
Original assignee: Leibniz-Institut für Polymerforschung Dresden e. V.
Priority date: 2016-04-04
Filing date: 2017-04-04
Publication date: 2017-10-12
Also published as: DE102016106097B3

Abstract

The invention relates to a tissue- and organ transport device (1) for transferring, transporting and stabilizing tissue and organs (6), comprising a rigid container (2) and a macroporous support material (4) for the tissue (6) or organ (6), said macroporous support material (4) being either securely or insecurely packed in the rigid container (2), having a pore size of ≥ 1 μm and having a system of interconnected pores. The tissue- and organ transport device according to the invention ensures that the arrangement of the tissue can be maintained and causes no damage, or does not lead to any damage to the tissue or organ during transport.

Description

Tissue and organ transport device

The invention relates to a tissue and organ transport device. 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 a growth medium. For transport from the tissue bank to the surgeon, the corneas are in chambers with a simple fastening system, which protects the tissue from swimming around during transport, set, for example, in a so-called Böhnke chamber with a corneal holder. In this connection, there is no transport system at all for precut endothelial lamellae (innermost single-layer of the cornea) of donor corneas, since they are very thin (100 μm). The fragile tissue must therefore remain in the bed of the donor cornea so that it can be transported from the tissue bank to the surgeon without damage.

JP 2002-335 950 A describes a container for storing and transporting a membrane tissue 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.

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.

JP 2010-220488 A describes an instrument with which cell culture products, such as cultivated tissue, can be taken up. The instrument consists of a plastic container with a porous wall, which when contacting the cell culture products, for example, this record using a controlled vacuum and release by an applied pressure. This instrument is intended to facilitate the handling of cells and their products, for example cell layers, in laboratories, but on the other hand does not allow storage or transport outside of a sterile environment. DE 10 2005 004 826 A1 describes a suction cup for the temporary removal of at least part 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 pm. As possible materials for the plant body various ceramic materials are specified.

From 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 a customized tissue and organ transport device which is suitable as a mechanical support for both 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 tissue and organ transport device must ensure sustained nutrient delivery 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 In addition, the organ transport device should permit 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 as well as a macroporous support material for the tissue or organ, the macroporous support material either embedded in the solid container or embedded unattached is. The invention is based on macroporous support materials having a minimum pore size of> 1 pm, preferably of> 2 μι, and with an interconnective pore system, that is, a system of interconnected pores. A carrier material formed with such a pore system enables a hydrodynamic flow. 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 stabilized, transported and transplanted / implanted. For example, the tissue and organ transport device may be in the form of a flat or tall cylinder or in the form of a syringe. The solid container preferably has a lid and a lower vessel, wherein the macroporous carrier material is at least embedded in the lower vessel, possibly also in the lid, either fixed or unpaved. Upon application of hydrodynamic pressure, for example via a syringe system, the tissue or organ may be carefully "pushed out" of the device and applied to the site of implantation in the body of the patient without the use of forceps and other surgical equipment In a preferred embodiment of the invention, the tissue and organ transport device comprises a solid container having a lid and a bottom vessel, wherein for the tissue or organ, the macroporous carrier material is embedded with the system of interconnected pores secured in the bottom vessel and allows hydrodynamic flow and Lower vessel is formed with a movable piston for generating a hydrodynamic pressure.

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 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 WL Murphy et al. 2002, Salt fusion: an approach to improve pore interconnectivity within tissue engineering scaffolds. Tissiue 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 TD Dziubla 2002 Macroporous Hydrogels as Vascularizable 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. Be. 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 material 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 the sustainable supply of nutrients and enables continuous supply with 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:

Fig. 1A: a schematic representation of an inventive

Tissue and organ transport device according to a first variant (macroporous support material only in the lower vessel), Fig. 1 B: a schematic representation of a tissue and

Organ transport device according to a second variant (macroporous support material both in the lower vessel and in the lid), Fig. 1C is a schematic representation of a tissue and

Organ transport device as a high cylinder with regions for generating a hydrodynamic pressure,

Fig. 1 D: a schematic representation of a tissue and

Organ transport device which is in the form of a syringe,

Fig. 2 shows an embodiment for an application of a macroporous support materials based, designed device for the recording and transmission of implantable tissues and organs and

3: a schematic representation of the transmission and the Transporting a delicate tissue using a syringe-shaped tissue and organ transport device.

1 A shows a first variant of the tissue and organ transport device 1 according to the invention, this device 1 comprising a solid container 2 with a lower vessel 3 in the form of a flat cylinder, in which a macroporous carrier material 4 is embedded. On the surface of the macroporous support material 4 in the middle of the cylindrical lower vessel 3, a cavity 5 for a tissue 6 or 6 organ is formed. Tissue 6 or organs 6 can be placed in or on the macroporous support material 4, which is embedded in a solid container 2. Furthermore, the solid container 2 according to FIG. 1A also has a cover 7 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 4 allows a gentle, gentle hydrodynamic flow. These two advantageous properties are indicated by the double arrow in the detail section of FIG. 1A.

1 B shows a second variant of the tissue and organ transport device 1, the device 1 also comprising a fixed container 2 with a lower vessel 3 in the form of a flat cylinder, in which a macroporous carrier material 4 is embedded. On the surface of the macroporous support material 4 in the middle of the cylindrical lower vessel 3, a cavity 5 for a tissue / organ 6 is formed. Tissues / organs 6 can thus be placed in or on the macroporous support material 4, which is embedded in a solid container 2. In contrast to FIG. 1A, in the variant according to FIG. 1B, porous carrier material 4 with a cavity 5 placed in the middle for the tissue / organ 6 is also embedded in the underside of the lid 7, which is in the form of a flat cylinder , As shown in Fig. 1 B, this cavity 5 is the macroporous Carrier material 4 in the lid 7 the cavity 5 of the substrate 4 in the lower vessel 3 exactly opposite. After closing the solid container 2, 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 envelop the fabric / organ 6 together as a united cavity. The advantage of the macroporous carrier material 4 is a cushioning effect for the container Tissue or organ. Furthermore, the porous carrier material 4 allows a gentle, gentle hydrodynamic flow. The advantages of using the macroporous support material 4 (cushioning effect and gentle hydrodynamic flow) are schematically indicated in the detail section of FIG. 1B by the double arrow.

FIG. 1C shows a third variant of the tissue and organ transport device 1. In this case, the tissue and organ transport device 1 comprises a fixed container 2, which has a cylindrical lower vessel 3 and a cover 7. In the underside of the lid 7, which is formed in the form of a flat cylinder, macroporous support material 4 is embedded with a placed in the center of the cylinder cavity 5 for the tissue / organ. The lower vessel 3 here is not formed in the form of a flat cylinder, but as a high cylinder, which has areas 8 for generating a hydrodynamic pressure below the carrier material 4 and the cavity 5, wherein in FIG. 1C the application of a hydrodynamic pressure in different directions is shown schematically by double arrows.

Finally, Fig. 1 D shows a fourth variant of the tissue and organ transport device 1, wherein also according to this variant in the bottom of the lid 7, which is in the form of a flat cylinder, macroporous carrier material 4 with a placed in the center of the cylinder cavity 5 is embedded for the tissue / organ 6. The lower vessel 3 'is in the form of a syringe 3' with a movable piston 9, which is movable via an operating element 10 in the cylindrical lower vessel 3 'in the axial direction is. By an axial movement of the piston, a hydrodynamic pressure can be generated along / parallel to the axial direction in the regions 8 which are located in the lower vessel 3 'below the carrier material 4 and the centrally placed cavity 5 with the tissue / organ 6.

The macroporous structure of the substrate 4 with 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 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 any 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 shown in FIG. 1D, the tissue or organ can be carefully removed from the device 1 without a great mechanical action, ie without the use of tweezers and other surgical devices "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 organ culture and growth media. The system of interconnected pores ensures the continuous and unobstructed supply of the tissue / organ. 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, this may contain chemical or biological entities functionalized to secure and support the viability and growth of tissues and organs to be transported and implanted. The applicability of the tissue and organ transport device for a very demanding task was investigated: the stabilization and transfer of a cellular monolayer generated by tissue engineering, as shown schematically in FIG. In this case, the cellular monolayer 11 is located on a thermoresponsive cell culture carrier 12, that is to say a carrier with a thermoresponsive polymer. Stimulable cell culture carriers 12, for example thermoresponsive cell culture carriers 12, are suitable for the culture of cellular monolayers 11 of human corneal endothelial cells. By using an external trigger, for example a decrease in temperature, detachment 13 of the cellular monolayer 11 from the stimulable cell culture carrier 12 takes place. This so-called cellular monolayer 11 is extremely thin (about 10 pm) and brittle and can therefore neither be touched nor lifted by forceps , It is known that cell turfs consisting 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 (so-called cryogels) as support material 14, detached endothelial cell layers 11 could be stabilized from human corneal endothelial cells and transferred to pigskin hulls as target substrates 15. 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, an aqueous gel-forming reaction mixture composed of starPEG and heparin was treated at temperatures of -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 survival of the transmitted human endothelial cell layers of the cornea. 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.

3 shows schematically the transmission and transport of the endothelium. Lamellae from the cornea as an example of a sensitive tissue using a syringe-like tissue and organ transport device shown in FIG. 1 D. It is carried out in step I) first gentle application of endothelial lamellae to the macroporous support material (gel) by generating a liquid flow through Moving the piston in the suction direction. The macroporous gel is relaxed and generates a slight negative pressure by swelling.

The transport takes place in method step II) while maintaining the orientation of the fabric due to existing 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 and thereby releases the liquid contained in the pores (for example cell culture medium) again.

LIST OF REFERENCE NUMBERS

1 tissue and organ transport device, device

2 solid containers

3 lower vessel

3 'lower jar in syringe form, syringe

4 macroporous carrier material

5 cavity in the substrate 4

6 organ, tissue

7 lids

8 areas for generating a hydrodynamic pressure

9 pistons

10 control element (for the piston 9)

11 cellular monolayer, endothelial cell layer

12 stimulable cell culture carrier

13 detachment of the cellular monolayer 11 from the stimulable cell culture carrier

14 biohybrid hydrogels

15 target substrate

Claims

claims

1. tissue and organ transport device (1) for the transmission, transport and stabilization of tissues and organs (6) comprising a solid container (2) and a macroporous support material (4) for the tissue (6) or the organ ( 6), wherein the macroporous support material (4) is embedded in the solid container (2) either fixed or unpaved, has a pore size of> 1 pm, and comprises a system of interconnected pores.

2. tissue and organ transport device (1) according to claim 1, characterized in that the solid container (2) has a cover (7) and a lower vessel (3, 3 '), wherein the macroporous support material (4) at least in the lower vessel ( 3, 3 ') is either fixed or embedded unattached.

3. 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) is adapted to the tissue to be transported (6) or organ (6).

4. 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) is adapted to the shape and geometry of the tissue to be transported (6) or organ (6).

5. tissue and organ transport device (1) according to one of claims 1 to 4, characterized in that the tissue and organ transport device (1) comprises a fixed container (2) with a lid and a lower vessel (3, 3 '), wherein for the tissue (6) or the Organ (6) the macroporous support material (4) is embedded with the system of interconnected pores in the lower vessel (3, 3 ') embedded and the lower vessel (3, 3') with a movable piston (9) for generating a hydrodynamic pressure is trained.

6. tissue and organ transport device (1) according to one of claims 1 to 5, characterized in that macroporous support material (4) is functionalized with chemical and / or biological entities.

7. tissue and organ transport device (1) according to one of claims 1 to 6, characterized in that the macroporous support material (4) is swellable by contact with an aqueous medium.

8. tissue and organ transport device (1) according to one of claims 1 to 7, characterized in that the macroporous support material (4) is prepared by a method selected from porogen leaching, solid-state foam formation, gas-foaming, phase separation, electrospinning and Kryogelbildung in an aqueous medium.

9. tissue and organ transport device (1) according to any one of claims 1 to 8 for use in the transplantation or implantation of tissues or organs.

10. Use of a tissue and organ transport device (1) according to any one of claims 1 to 9 for the transmission, transport and stabilization of transplanted or implanted tissues (6) or organs (6).