METHOD TO PACKAGE A TISSUE MATRIX TO BE REGENERATED

Abstract

Biological tissue is packaged to be regenerated before grafting in a vial sealed under vacuum and comprising a biological tissue matrix. A method for producing said vial includes placing a treated ex-vivo tissue sample in an open rigid vial and placing the vial in a lyophilizer. A lyophilization process is performed under vacuum to convert the treated ex-vivo tissue sample into a biological tissue matrix. The vial is hermetically sealed with closing means, inside the lyophilizer under vacuum. The sealed vial is then removed from the lyophilizer.

Claims

1. A method for producing a vial, under vacuum, including a biological tissue matrix to be regenerated for grafting, the method comprising: placing a treated ex-vivo tissue sample in an open rigid vial; placing the vial in a lyophilizer; running a lyophilization process under vacuum to convert the treated ex-vivo tissue sample into a biological tissue matrix; hermetically sealing the vial with closing means, inside the lyophilizer under vacuum, and removing the sealed vial from the lyophilizer.

2. The method according to claim 1, further comprising, before running the lyophilization process: pre-positioning the closing means onto the vial in a manner enabling fluid communication through the opening of the vial.

3. The method according to claim 1, wherein hermetically sealing the vial with the closing means comprises applying a pressure onto the closing means.

4. The method according to claim 1, performed under sterile conditions.

5. A vial sealed under vacuum with closing means and comprising a biological tissue matrix.

6. The vial according to claim 5, wherein the closing means comprise a rubber stopper or an injectable membrane.

7. The vial according to claim 5, wherein the closing means is suitable for reversible needle perforation.

8. The vial according to claim 5, wherein the vial is made of glass or rigid polymer.

9. The vial according to claim 5 wherein the tissue matrix has a rate of humidity below 10% by weight, preferably below 6% by weight.

10. The vial according to claim 5, wherein the tissue matrix is sterile and is for pharmaceutical use, clinical use or veterinary use.

11. A use of the vial of claim 4 for producing a regenerated tissue from the biological tissue matrix.

12. The method to regenerate a biological tissue matrix packaged in a vial according to claim 5, comprising the steps of: introducing in the vial a regenerating fluid to regenerate the tissue matrix to a biological tissue.

13. The method according to claim 10, wherein introducing the regenerating fluid in the vial comprises injecting, inside the vial, regenerating fluid contained in a syringe by: reversibly perforating the opening means of the vial with the needle of the syringe; transferring the regenerating fluid from the syringe to the vial, and withdrawing the needle of the syringe from the opening means.

14. The method according to claim 10, wherein the regenerating fluid comprises cells in suspension in blood, plasma rich platelet, bone marrow, lipoaspirate, a growth factors solution, a culture media with stem cells or a suspension of a bioactive agent.

15. The method according to claim 1, wherein the tissue matrix originates from one of: bone, skin, dermis, tendon, cortical bone powder, bladder, and skeletal muscle

Description

[0035] The invention will be better understood with the following description of several examples, referring to the accompanying drawing on which:

[0036] FIG. 1 illustrates the method of the invention.

[0037] Referring to FIG. 1, an ex-vivo tissue sample, which originates from an biological tissue having undergone several cleaning steps, manually, for example as disclosed in Biomaterials 23 (2002) 2979-2988, or in an automated manner, as disclosed in PCT/EP2017/083540, is introduced in a vial 2 in a step A. The ex-vivo tissue sample 1 can be here, for example, a piece of bone matrix, and the vial 2 is here a cylindrical glass vial, with a narrower neck 3.

[0038] In a step B, a stopper 4, here in rubber, is positioned partially inserted in the neck 3 of the vial 2, in such a way that an opening 5 remains for fluid communication between the inside and the outside of the vial.

[0039] This step B of pre-positioning the stopper on the opening of the vial is particularly adapted for the kind of stopper 4 illustrated, having legs 14 enabling the stopper to be blocked in the neck of the vial while leaving an open space 5 between the legs 14 to enable circulation of gas and humidity between the ex-vivo tissue sample 1 and the outside of the vial 2.

[0040] However, there are other means to insert the stopper in the neck of the vial at later stages of the method, as will become apparent below.

[0041] In step C, the vial 2 containing the ex-vivo tissue sample 1 and topped with stopper 4 in semi-open position in placed in the lyophilizer 6. A simple lyophilizer containing three vials is here illustrated for simplicity, but a lyophilizer can usually contain several vials, even several hundreds of vials, depending on their size, laid out on one or several shelves designed to accommodate vials in a way that they cannot freely move around on the shelf (for example with a rack), or under other arrangements depending on the configuration of the lyophilizer.

[0042] When all vials to be lyophilized have been introduced in the lyophilizer, the lyophilizer is closed and the lyophilization process can be performed in step D. Typically, the temperature inside the lyophilizer is reduced to freeze the content of the vials, here the ex-vivo tissue sample 1, and the pressure is reduced to the point of sublimation of the water content of the ex-vivo tissue sample 1.

[0043] The process is continued to obtain the tissue matrix 10, as dry as possible, preferably a water content below 10% by weight and still preferably of 6% by weight or less.

[0044] At the end of the lyophilization process, in step E, before the lyophilizer is open and the vacuum breached, the stoppers 4 are pressed upon for their complete insertion into the neck 3 of the vial. That way, the tissue matrix 10, almost completely dehydrated, is isolated from the outside of the vial, under vacuum.

[0045] In practice, an actionable plate 7 can be installed inside the lyophilizer, above the vials. At the end of the lyophilization process, the plate 7 can be actioned downward in order to exert a pressure on the pre-positioned stopper(s) 4, until the stoppers are completely inserted in the neck 3 of the vial(s) 2.

[0046] Alternatively, the stopper(s) can be initially attached/adhered to the plate 7 and be inserted fixedly in the neck 3 of the vials 2 upon a downward pression. It could also be envisaged, when an array of vials is arranged on a shelf of a lyophilizer, that the plate or a roll arranged in the lyophilizer deposits a film on the vials, and that the film is thermo-sealed to the top collar of the neck, by application of heat coming, for example from the plate or the same roll, which could be heatable.

[0047] The person skilled in the art thereby understands that there are several technical manners to implement a step of sealing the vials inside a lyophilizer.

[0048] In step F, the lyophilizer is open and the vial 2, sealed, and comprising a dehydrated biological tissue matrix 10 under vacuum, to be regenerated for grafting, is removed.

[0049] The matrix 10 is now packaged in a form that will allow a long shelf life and an easy use. The vacuum inside the vial will have a tendency to exert a suction onto the rubber stopper, thereby helping maintaining it in place. Because the tissue matrix is not exposed to ambient air after lyophilization, the humidity content of the packaged material remains below 10% by weight and preferably below 6% by weight, allowing a much longer shelf—life of the tissue matrix compared to existing matrixes currently on the market.

[0050] Optionally, to reinforce the sealing of the vial, in particular for rubber stoppers and glass vials, an aluminum cap 8 can be placed on the sealed vial 2 in step G, and the cap can be crimped onto the vial neck 3 in step F, in order to fasten the cap 8 and rubber stopper 4 to the vial and avoid any unwanted opening.

[0051] Advantageously the cap does not fully cover the top of the stopper or comprises an openable strip to easily uncover an area of the top of the rubber stopper. This present some advantages for further use of the tissue matrix in the vial.

[0052] Though a single cubic piece of bone matrix 10 is here illustrated in the vial 2, the invention is not limited to any particular tissue matrix to be regenerated, having any kind of shape. It could for example be other tissue matrixes like skin, dermis, tendon, cortical bone powder, bladder, skeletal muscle . . . .

[0053] There may be, in certain cases more than one piece of tissue matrix per vial.

[0054] A step of labeling of the vial can be foreseen, in case labeling was not performed before. The label typically contains an identifier (barcode, QR code, RFID chip . . . ) for traceability of the tissue matrix, as is well known in the field.

[0055] The tissue matrix can have been prepared through cleaning steps performed by an operator in a sterile facility, in a completely manual way or using some automated steps. The tissue matrix can also have been prepared in a completely automated manner, like for example in a reactor as described in PCT/EP2017/083540 which comprises: [0056] an airlock entry for introduction and classification (introduction into sterile area responding to regulatory standards) of a biological tissue; [0057] means to treat the biological tissue arranged to produce a tissue matrix; [0058] means for packaging the tissue matrix; [0059] an airlock exit of the packaged tissue matrix; [0060] automated means to move the biological tissue from the airlock entry to the treatment means, and [0061] automated means to move the biological tissue matrix from the treatments means to the packaging means and further to the airlock exit.

[0062] In such a case, the method of the invention is related to the packaging means as well as the treatment means (lyophilisation).

[0063] The method to regenerate the biological tissue matrix 10, packaged in the vial 2 sealed under vacuum, comprises introducing in the vial a regenerating fluid to regenerate the tissue matrix to a biological tissue. After the tissue is regenerated, it will suitable to be grafted to a patient.

[0064] Referring to FIG. 2, the step I of introducing in the vial a regenerating fluid to regenerate the tissue matrix 10 to a biological tissue can be performed through the use of a syringe initially containing the regenerating fluid 21. The free extremity of the needle 22 of the syringe 20 is introduced in the vial 2 through the stopper 4, which can be a rubber stopper or an injectable membrane. Through pressing on the piston of the syringe and/or suction force resulting from the pressure difference between the inside of the vial and the external environment, the regeneration fluid is transferred into the vial, onto the tissue matrix 10. The needle of the syringe is then removed from the stopper which re-seals, leaving no opening (hence the reversible perforation of the stopper by the needle).

[0065] After a period of time, which can depend on the type of tissue, the matrix is regenerated to a biological tissue, suitable to be grafted to a patient. The professional merely needs to open the vial and sample the tissue.

[0066] The regenerating fluid can be cells in suspension in blood, plasma rich platelet, bone marrow, lipoaspirate, a growth factors solution, a culture media with stem cells or a suspension of a bioactive agent . . . .

[0067] The advantage of using the tissue matrix in the vial as described above is that sterility of the tissue can be ensured up to the last minute before grafting, as the number of manipulation step of the tissue is reduced, the tissue is not put in contact with other containers and the vial remains has closed to any potential contamination.

[0068] Alternatively, in case the stopper is not suitable for a needle perforation, for example when the tissue matrix is packaged under vacuum in a plastic tray sealed with a plastic film as closing means, the film is removed, regeneration fluid is added in the vial. Time is allowed for regeneration of the tissue matrix to a biological tissue suitable for grafting. These steps can be performed directly in the operating theatre, where contamination is highly controlled.

[0069] The regenerating fluid can originate from the patient himself and can for example be cells in suspension in blood, plasma rich platelet, bone marrow, lipoaspirate, a growth factors solution, a culture media with stem cells or any other bioactive agents. The tissue matrix can originate from the patient himself and can, for example, be bone, skin, dermis, tendon, cortical bone powder, bladder, skeletal muscle.

[0070] A patient can be a human patient or an animal.