Multifunctional device, device holder, kit, use of bioactive factors isolated and purified from colostrum, method for preparing enriched autologous platelet concentrates

Abstract

The present invention relates to a multifunctional device (1) for collecting a biological fluid, for example whole blood, comprising: a tubular container (2) delimiting an open container end (3), a cap (5), wherein the cap (5) is fluid-tightly connected to the tubular container (2), closing the open container end (3) so that the tubular container (2) and the cap (5) enclose a collection chamber (4) configured to be depressurized and / or filled with the biological fluid, wherein the cap (5) comprises: a cap body (6) connected to the tubular fluid-tight container (2), a reversible access device (7), wherein the reversible access device (7) is fluid-tightly connected to the cap body (6), wherein the reversible access device (7) is configured to allow a reversible fluid-tight access to the collection chamber (4) without separating the cap (5) from the tubular container (2) to depressurize the collection chamber and / or collect the biological fluid in the collection chamber (4) and / or extract the biological fluid from the collection chamber (4), the cap (5) comprising at least one reservoir (8, 8') fluid-tightly connected to the cap body (6), wherein each reservoir (8, 8') delimits a respective containment chamber (9) configured to contain at least one mixing substance, wherein each reservoir (8, 8') is in a closed configuration in which the respective containment chamber (9) is fluidly isolated from the collection chamber (4), wherein each reservoir (8, 8') is configured to be operated and opened on command, switching from the closed configuration to an open configuration in which the respective containment chamber (9) is open and in communication with the collection chamber (4) keeping the cap (5) fluid-tightly connected to the tubular container (2).

Description

Multifunctional device, device holder, kit, use of bioactive factors isolated and purified from colostrum, method for preparing enriched autologous platelet concentratesDESCRIPTION

[0001] . Field of the inventionThe present invention relates to a multifunctional device for collecting a biological fluid, as well as a device holder, a kit, a use of bioactive factors isolated and purified from colostrum, and a method for preparing autologous platelet concentrates (Platelet-Rich Plasma or PRP or platelet gel, Leukocyte- Rich PRP or L-PRP, Plasma Rich in Growth Factors or PRGF, Platelet-Poor Plasma or PPP, etc.) enriched with bioactive factors isolated and purified from colostrum, to obtain, in a single closed, sterile and vacuum environment, a (liquid or gel) product with improved bioactive properties for use in treating conditions requiring tissue repair and regeneration, in humans and animals.

[0002] . Background art

[0003] . Tissue injury healing is a complex process involving a cascade of cellular and molecular events mostly shared by the different tissues of the body. Interestingly, the tissue repair process begins immediately after a traumatic injury and is mediated and controlled by a wide range of cytokines, proteins, and growth factors, some of which are released by platelets upon activation.

[0004] . In recent years, the development of platelet-rich preparations has revolutionized the field of regenerative medicine due to the repair capacity of growth factors, released by the platelets themselves, which stimulate and accelerate tissue healing and regeneration.

[0005] . Today, autologous platelet concentrates (APCs) are widely used in a wide range of medical disciplines which require rapid tissue healing and regeneration.

[0006] . APCs are blood components obtained by centrifuging the patient’s blood in order to collect the most active components: platelets, fibrin and, in some cases, even leukocytes. The final product has a platelet concentration above the baseline level, consequently it has a higher number of growth factors derived from the platelets themselves. The rationale for the clinical use of such preparations is based on the concept of exploiting the enriched content thereof of several platelet- derived mitogenic growth factors (including Platelet-Derived Growth Factor or PDGF, Transforming Growth Factor-p or TGF-p, Endothelial Growth Factor or EGF, Vascular Endothelial Growth Factor or VEGF, Insulin-like Growth Factor-1 or IGF-1 , basic Fibroblast Growth Factor or bFGF and Hepatocyte Growth Factor or HGF) to stimulate various biological functions, such as chemotaxis, angiogenesis, proliferation and differentiation, so as to promote the healing of hard and soft tissues.

[0007] . The term “platelet-rich plasma” (PRP) was first introduced by Kingsley et al. to describe a platelet concentrate used for the treatment of severe thrombocytopenia. However, the use of theterm PRP really began with Marx in 1998 when he published a comparative clinical study in which the regenerative potential of PRP was demonstrated in a series of patients undergoing mandibular reconstruction. PRP was then associated with the concept of platelet growth factors and the potential contribution thereof in inducing tissue healing.

[0008] . There are currently several commercial systems for preparing PRP which can lead to products with different features, in particular related to the composition and the cellular concentration rate with respect to the baseline. On average, a 5-8x concentration is obtained, although a ratio of up to 11x has been reported with PRP.

[0009] . Although the form thereof can vary based on different applications (liquid form, gel, membranes or fibrin clots, etc.), as well as the preparation protocols, there is multiple clinical evidence on the efficacy of autologous platelet concentrates (APCs) in many fields of medicine, the fundamental role thereof in the regeneration of different tissues having been demonstrated: bones, skin, oral mucosa, tendons, ligaments, muscles, cartilage, cornea, etc., thereby comprising all medical specialties, such as oral and maxillofacial surgery, orthopedics and sports medicine, dermatology and aesthetic medicine, gynecology, andrology, plastic and reconstructive surgery, ophthalmology, otolaryngology, neuroscience, internal medicine (organ injuries) and even veterinary medicine and surgery.

[0010] . However, as can be seen from the several studies reported in the literature, for many applications of Autologous Platelet Concentrates (APCs) the results obtained were not satisfactory, and even for those applications for which the effects of treatment with APCs have been positively judged, the results obtained are clearly susceptible to further notable improvements.

[0011] . Therefore, an attempt was made to enhance the effect of APCs by using them in combination with adult stem cells taken from the same patient, antibiotics, anti-inflammatory agents, vitamins, etc., however the increase in efficacy obtained with these combinations, with respect to the use of APCs alone, was quite modest.

[0012] . In WO2022079684A1 it has been disclosed that by combining biological factors extracted from colostrum in active form with Autologous Platelet Concentrates (APCs), the activity of the latter is significantly enhanced in all application fields thereof. By combining the APCs with the bioactive factors derived from colostrum, the two main defects of the APCs are solved, i.e., the lack of some fundamental factors to modulate a correct growth and regeneration of the tissues and the lack of substances which provide the indispensable nourishment to the cells being grown and proliferating.

[0013] . According to the protocol for the production of PRP, depicted in Figure 16, the patient’s blood is collected in test tubes containing anticoagulants and processed by means of two centrifugation steps. The PRP thus obtained can be applied to the site to be treated with a syringe or activated by thrombin and / or calcium chloride to trigger platelet activation and stimulate fibrinpolymerization.

[0014] . After the collection of blood in test tubes with anticoagulant, a first low-intensity centrifugation (soft spin) allows the separation of the blood into three distinct layers: red blood cells at the bottom, a cell plasma (Platelet-Poor Plasma or PPP) at the top and a whitish layer referred to as the buffy coat, located therebetween, containing the highest concentration of platelets and leukocytes. For the production of Pure-PRP (P-PRP), the PPP and the buffy coat surface layer are transferred to another test tube and centrifuged at high intensity (hard spin), after which most of the PPP and leukocytes are discarded and the P-PRP can be collected. To obtain the leukocyte rich PRP (L-PRP), the PPP, the entire buffy coat layer and some residual red blood cells are collected and transferred to another test tube to be centrifuged in a second centrifugation at high intensity (hard spin), after which most of the PPP is discarded thus obtaining an L-PRP containing the buffy coat with most of the platelets and leukocytes, some residual red blood cells and PPP.

[0015] . The PRP production protocol can also include activating the APCs by mixing with particular substances (e.g., 0.2 mL of 10% CaCh, with or without thrombin) which cause platelet degranulation (i.e., the release of the growth factors contained in the platelets themselves) and the immediate formation of the platelet gel, substances which are added to the obtained PRP or which are arranged in a further tube to which the PRP is added.

[0016] . The PRP production protocol can also include mixing the autologous platelet concentrates with further substances, such as with bioactive factors derived from colostrum, making it necessary to transfer the APCs into other tubes with the substances to be mixed or vice versa.

[0017] . During the first centrifugation, the second centrifugation, and the mixing step of the autologous platelet concentrates, with the opening of the tubes for the transfer of the content thereof into a further tube or for the addition of substances to be mixed, the final product is exposed to the risk of infectious contamination and loss of sterility which can compromise the use of the product. For example, in WO2013 / 111130 it is expected to collect whole blood in a tube comprising a container closed by a cap, centrifuge the tube to separate the whole blood, and after centrifuging the tube, insert a needle into the tube and use a syringe to withdraw a portion of the whole blood separated from the tube to mix it with a mixing substance contained in the syringe, where the transfer of the contents of the tube into the syringe to be mixed introduces a risk of error in the mixing step and a risk of infectious contamination and loss of sterility which can compromise the use of the product, and where the mixing is carried out in a container different from the tube in which the whole blood was collected.

[0018] . The need is therefore strongly felt in the field to create solutions of collection devices for biological fluids to then be mixed, which allow avoiding infectious contamination of the contents of the collection device.

[0019] . Solution

[0020] . These and other objects are achieved by a multifunctional device, a device holder, a kit, a method for preparing autologous platelet concentrates enriched with bioactive factors isolated and purified from colostrum, as well as a use of bioactive factors isolated and purified from colostrum in the preparation of enriched autologous platelet concentrates and simultaneously to cause platelet degranulation and platelet clot formation.

[0021] . Some advantageous embodiments are the subject of the dependent claims.

[0022] . According to an aspect, the present invention allows sampling whole blood, separating red blood cells from white blood cells, plasma and platelets (PPP and PRP), mixing, under aseptic conditions, the latter fraction with the bioactive factors isolated and purified from colostrum and the simultaneous activation of the enriched PRP thus formed (or of the other APCs).

[0023] . According to an aspect, the present invention allows obtaining a platelet clot, enriched with bioactive factors derived from colostrum, without resorting to the use of further platelet activators.

[0024] . According to one an , the present invention allows preparing Autologous Platelet Concentrates (APCs) activated and enriched with bioactive factors isolated and purified from colostrum, and possibly further substances.

[0025] . By virtue of the suggested solutions, it is possible to avoid the frequent movements of blood / plasma / APCs between syringes and tubes and vice versa, thus allowing activated Enriched Autologous Platelet Concentrates (EAPCs) to be obtained, which are totally sterile and free of any environmental pollutants.

[0026] . By virtue of the suggested solutions, it is possible to make a multifunctional device which allows sampling a biological fluid, such as whole blood, the separation thereof into part liquid (plasma) and part particulate (red blood cells, white blood cells and platelets) and mixing, under aseptic conditions, the PRP and PPP thus obtained with the bioactive factors isolated and purified from colostrum, contained therein.

[0027] . By virtue of the suggested solutions, it is possible to obtain a blood sampling or collection, a separation thereof into fractions and an integration of the growth factors present in the PRP and PPP with those derived from colostrum, all in a single environment (the multifunctional device) which is closed, sterile and vacuum, so as to achieve the goals of extreme safety and high efficiency.

[0028] . By virtue of the suggested solutions, it is surprisingly possible to use bioactive factors isolated and purified from colostrum to activate platelets, and obtain the platelet clot without resorting to the use of calcium chloride (typically used in amounts of about 0.2 mL of 10% CaCh, with or without thrombin), for platelet activation.Furthermore, the suggested solution allows always operating within a single closed, sterile and vacuum environment (the multifunctional device).

[0029] . Figures

[0030] . Further features and advantages of the invention will become apparent from the description provided below of preferred embodiments thereof, given by way of non-limiting indication, with reference to the accompanying drawings, in which:

[0031] . - Figure 1 depicts a whole blood collection kit according to the prior art comprising a tube provided with a cap comprising a rubber body configured to be pierced to collect blood and a cap configured to be connected to the tube container, locking the rubber body of the cap in place, a tube holder or sleeve configured to accommodate the tube and pierce the membrane of the tube, in which the tube holder has a central hole, preferably a Luer hole, connectable on one side to an adapter, preferably a Luer adapter, connected to an inlet needle, e.g., a butterfly needle, for sampling blood by means of a sampling tube, and on the other side an outlet needle configured to be connected to the tube holder and to pierce the tube cap and transfer the blood from the butterfly needle to the tube;

[0032] . - Figure 2 shows an axonometric view of a kit according to the present invention, comprising a sampling assembly, comprising an inlet needle, preferably butterfly, connected by means of a sampling tube to an outlet needle provided with a connection adapter, preferably a Luer adapter, a device holder or sleeve to which the outlet needle is connectable, and a multifunctional device according to a first embodiment, in which the multifunctional device comprises a cap comprising a reversible access device, e.g., a cap portion configured to be pierced to insert or withdraw the contents of the multifunctional device, in which the cap comprises at least one reservoir to contain at least one substance to be mixed on command with the contents collected in the tube without having to separate the cap from the tubular container of the multifunctional device, avoiding the contents collected in the multifunctional device from being exposed to the external environment;

[0033] . - Figure 3 depicts an axonometric view of a multifunctional device according to a first embodiment used in the kit in Figure 2;

[0034] . - Figure 4 shows a multifunctional device according to a second embodiment usable in the kit in Figure 2, in which the cap comprises at least two reservoirs to contain each at least one substance to be mixed on command with the collected blood;

[0035] . - Figure 5 depicts a first side view of the multifunctional device in Figure 3;

[0036] . - Figure 6 depicts a second side view of the multifunctional device in Figure 3;

[0037] . - Figure 7 shows a top view of the multifunctional device in Figure 3;

[0038] . - Figure 8 shows a first side view of the multifunctional device in Figure 4;

[0039] . - Figure 9 shows a second side view of the multifunctional device in Figure 4;

[0040] . - Figure 10 shows a top view of the multifunctional device in Figure 4;

[0041] . - Figure 11 shows an axonometric view of a device holder or sleeve, preferably bellshaped, configured to accommodate a multifunctional device according to the present invention, inwhich the device holder or sleeve comprises a sampling hole for a connection to a sampling tube and to an outlet needle, and at least one opening configured for the passage without interference of the at least one reservoir of the cap of the multifunctional device, from which the reservoir of the cap of the multifunctional device is arranged passing through;

[0042] . - Figure 12 shows a top view of the device holder in Figure 11 ;

[0043] . - Figure 13 shows a side view of the device holder in Figure 11 ;

[0044] . - Figure 14 shows an axonometric view of a sampling tube assembly comprising a sampling needle, or inlet needle, connected by means of a sampling tube to an adapter configured to be connected to the device holder, and an outlet needle, covered by a rubber protection, configured to be connected to the reversible access device of the cap of the multifunctional device for transferring whole blood into the multifunctional device according to the present invention;

[0045] . - Figure 15 is a table showing the growth factors derived from platelets and the functions thereof;

[0046] . - Figure 16 diagrammatically shows a protocol for producing PRP according to the prior art, in which one or more transfers between tubes of the centrifuged and / or mixed blood are involved, to obtain the final product;

[0047] . - Figure 17 depicts a flow diagram for the exosome purification procedure according to the prior art.

[0048] . Description of some preferred embodiments

[0049] . In accordance with a general embodiment, a multifunctional device for collecting a biological fluid, e.g., whole blood, is indicated by reference numeral 1.

[0050] . The multifunctional device 1 comprises a tubular container 2 delimiting an open container end 3. The multifunctional device 1 further comprises a cap 5. In an embodiment, the tubular container 2 comprises a curved or rounded bottom wall, opposite to the open container end 3. In an embodiment, the tubular container 2 comprises a side wall, preferably cylindrical, extending axially between a bottom wall and the open container end 3.

[0051] . The cap 5 is fluid-tightly connected to the tubular container 2, closing the open container end 3 so that the tubular container 2 and the cap 5 enclose a collection chamber 4 configured to be depressurized and / or filled with the biological fluid. In an embodiment, the multifunctional device 1 is configured to contain from 10 mL to 18 mL, preferably 12 mL, of whole blood in the collection chamber 4.

[0052] . The cap 5 comprises a cap body 6 fluid-tightly connected to the tubular container 2.

[0053] . The cap 5 further comprises a reversible access device 7. The reversible access device7 is fluid-tightly connected to the cap body 6. The reversible access device 7 is configured to allow a reversible fluid-tight access to the collection chamber 4 without separating the cap 5 from the tubularcontainer 2 to depressurize the collection chamber and / or to collect the biological fluid in the collection chamber 4 and / or to extract the biological fluid from the collection chamber 4. In accordance with an embodiment, the reversible access device 7 is a membrane made of fluid-tight elastomeric material, e.g., rubber, configured to be reversibly pierced by a needle, and once the needle is removed the membrane self-seals, fluidly isolating the collection chamber 4 from the external environment. In still other embodiments, the reversible access device 7 can be a valve which is normally closed by fluidly isolating the collection chamber from the external environment, and which can be opened by a user to depressurize the collection chamber or to insert the fluid into the collection chamber or to withdraw the contents of the collection chamber. In an embodiment, the cap body 6 comprises, in a single structure preferably made of fluid-tight elastomeric material, e.g., pre-molded soft rubber, the reversible access device 7.

[0054] . Advantageously, the cap 5 comprises at least one reservoir 8, 8’ fluid-tightly connected to the cap body 6. In an embodiment, the at least one reservoir 8, 8’ is integral with the cap body 6. In an embodiment, the at least one reservoir 8, 8’ is rigidly connected to the cap body 6, so as to be integral with the cap body 6.

[0055] . Each reservoir 8, 8’ delimits a respective containment chamber 9 configured to contain at least one mixing substance. In an embodiment, the reservoir contains from 50 mg to 250 mg, preferably 100 mg, of the at least one mixing substance, e.g., of bioactive factors isolated and purified from colostrum, and possibly other substances. In an embodiment, such a reservoir is preferably made of rigid plastic.

[0056] . Each reservoir 8, 8’ is in a closed configuration in which the respective containment chamber 9 is closed and fluidly isolated from the collection chamber 4, preventing a passage of the at least one mixing substance into the collection chamber 4.

[0057] . Each reservoir 8, 8’ is configured to be operated and opened on command, passing from the closed configuration to an open configuration in which the respective containment chamber 9 is open and in communication, e.g., in fluid communication or at least in fluid communication, with the collection chamber 4, allowing the passage of the at least one mixing substance in the collection chamber 4, without losing the fluid-tight connection between the cap 5 and the tubular container 2 of the multifunctional device 1. In an embodiment, each reservoir 8, 8’ in the open configuration is configured to release the mixing substance into the collection chamber 4. In an embodiment, each reservoir 8, 8’ in the open configuration is configured to release the mixing substance into the collection chamber 4 by gravity. In an embodiment, each reservoir 8, 8’ is configured to be operated and opened on command, passing from the closed configuration to the irreversibly open configuration, keeping the respective containment chamber 9 open and in communication with the collection chamber 4, and without losing the fluid-tight connection between the cap 5 and the tubular container 2of the multifunctional device 1. In an embodiment, each reservoir 8, 8’ is configured to be operated and opened on command, passing from the closed configuration to the open configuration, without accessing the collection chamber by means of the reversible access device 7.

[0058] . By virtue of the provision of a cap 5 comprising at least one reservoir 8, 8’ openable on command, keeping the cap 5 of the multifunctional device 1 fluid-tightly connected to the tubular container 2 of the multifunctional device 1 , it is possible to mix the biological fluid collected in the collection chamber 4 with the at least one mixing substance, previously stored in the reservoir 8, 8’ and fluidly isolated from the collection chamber 4, without exposing the collection chamber 4 to the external environment and without losing the fluid tightness between the cap 5 and the tubular container 2, and consequently avoiding any possible contamination of the contents of the multifunctional device 1 . Furthermore, by virtue of the provision of a cap 5 comprising said at least one reservoir 8, 8’, it is possible to mix the biological fluid collected in the collection chamber 4 with the at least one mixing substance previously stored in the reservoir 8, 8’, keeping the biological fluid collected in the collection chamber 4, without withdrawing a portion of biological fluid collected from the collection chamber 4 with a collection syringe, e.g., without piercing the cap with a needle of the collection syringe, to mix it with a mixing substance contained in the collection syringe, avoiding any possible contamination of the contents of the multifunctional device 1 . Furthermore, by virtue of the provision of a cap 5 comprising said at least one reservoir 8, 8’ it is possible to introduce into the collection chamber 4 the mixing substance previously stored in the reservoir 8, 8’ to mix it with the biological fluid contained in the collection chamber 4, avoiding withdrawing the biological fluid for mixing outside the multifunctional device. Furthermore, by virtue of the provision of a cap 5 comprising said at least one reservoir 8, 8’, it is possible to centrifuge the multifunctional device and then open the reservoir 8, 8’ on command to introduce the mixing substance into the collection chamber 4 to mix it with the collected biological fluid, avoiding putting the collection chamber 4 in communication, e.g., in fluid communication, with the external environment and with other collection devices unrelated to the multifunctional device 1 to mix the collected biological fluid.

[0059] . In accordance with an embodiment, the multifunctional device 1 comprises a safety cap 31 configured to be connected to the tubular container 2, and configured to form an axial abutment to avoid an undesired separation of the cap 5 from the tubular body 2. In accordance with an embodiment, the safety cap 31 is connectable and / or connected to the tubular container 2 by threaded connection. In accordance with an embodiment, the cap 5 comprises the safety cap 31 , in which the safety cap 31 is configured to avoid an undesired separation of the cap body 6, to which the reversible access device 7 and the at least one reservoir 8, 8’ are connected, from the tubular body 2, leaving free access to the reversible access device 7 and to the activation by command of the at least one reservoir 8, 8’. In accordance with an embodiment, the safety cap 31 is configured to avoid anundesired separation of the cap body 6 comprising, in a single structure or in one piece, the reversible access device 7 and the housing of the at least one reservoir. In accordance with an embodiment, the safety cap 31 is made of rigid plastic. In an embodiment, the safety cap 31 has a tubular shape and comprises at one end an axial retaining crown configured to abut against the cap body 6 and / or a portion of the cap 5.

[0060] . In an embodiment, each reservoir 8, 8’ delimits a respective open reservoir end 11.

[0061] . In an embodiment, the cap 5 comprises at least one separation device 10.

[0062] . Said at least one separation device 10, in said closed configuration of said reservoir 8,8’, is fluid-tightly connected to the reservoir 8, 8’, closing the respective open reservoir end 11 and fluidly isolating the respective containment chamber 9 from the collection chamber 4.

[0063] . Said at least one separation device 10, in said open configuration of said reservoir 8, 8’, is configured to delimit at least one passageway 12, putting the respective containment chamber 9 in communication with the collection chamber 4.

[0064] . Said at least one separation device 10 is operable on command, forming said at least one passageway 12 so as to switch on command the respective reservoir 8’, 8 from the closed configuration to the open configuration keeping the cap 5 fluid-tightly connected to the tubular container 2.

[0065] . In an embodiment, the cap body 6 delimits a reservoir housing 15 in which the reservoir 8, 8’ is housed and supported. The at least one reservoir 8, 8’ is connected to the cap body 6 so that there are no sealing leaks between the collection chamber 4 and the environment outside the multifunctional device 1 . For example, a sealing connection between the reservoir and the cap body 6 can be made from the material with which the cap 6 is made and / or by the provision of sealing gaskets between the outer walls of the reservoir and the walls of the cap body 6 delimiting the reservoir housing 15. In an embodiment, the cap body 6 comprises, in a single structure preferably made of fluid-tight elastomeric material, e.g., pre-molded soft rubber, the reversible access device 7 and the reservoir housing 15 in which the reservoir 8, 8’ is housed and supported.

[0066] . In an embodiment, the separation device 10 is fluid-tightly connected to the cap body 6 and delimits the reservoir housing 15 in the direction of the collection chamber 4. In an embodiment, the separation device 10 is that part of the cap body 6 delimiting the reservoir housing 15 in the direction of the collection chamber 4.

[0067] . In an embodiment, the reservoir 8, 8’, under an external thrust action, is configured to advance in the reservoir housing 15 in the direction of the collection chamber 4 from a resting position to at least an advanced position.

[0068] . In said resting position, the reservoir 8, 8’ is in said closed configuration of said reservoir

[0069] . In said at least one advanced position, the reservoir 8, 8’ is in said open configuration of said reservoir 8, 8’ in which the reservoir 8, 8’ operates the separation device 10 forming said at least one passageway 12.

[0070] . In an embodiment, the separation device 10 comprises a closing element, e.g., a hatch, connected to an opening, for example, of the cap body 6 or of the reservoir 8, 8’, which opening is normally closed by the fluid-tight closing element with respect to the reservoir 8, 8’ and with respect to the cap body 6, and which upon an advancement of the reservoir 8, 8’ or for the operation of another mechanism, the closing element opens, e.g., in the direction of the collection chamber 4, allowing the passage of the at least one mixing substance, in liquid and / or gel and / or solid form, in the collection chamber 4.

[0071] . In an embodiment, said at least one separation device 10 comprises at least one separation septum 13 constrained to the cap body 6.

[0072] . In an embodiment, the at least one separation septum 13 is fluid-tightly connected to the reservoir 8, 8’ in said closed reservoir configuration, and in which the at least one separation septum 13 is pierceable on command, forming said at least one passageway 12 so as to switch on command the respective reservoir 8, 8’ from the closed configuration to the open configuration, keeping the cap 5 fluid-tightly connected to the tubular container 2.

[0073] . In an embodiment, each reservoir 8, 8’ comprises a piercing portion 14 at least partially delimiting the respective open reservoir end 11. In an embodiment, the piercing portion 14 is a lance portion.

[0074] . In an embodiment, the piercing portion 14 is configured to pierce the at least one separation septum 13 following said external thrust action on the at least one reservoir 8, 8’ against the at least one separation septum 13 in the direction of the collection chamber 4, keeping the cap 5 fluid-tightly connected to the tubular container 2 and keeping the reservoir 8, 8’ fluid-tightly connected to the cap body 6.

[0075] . In an embodiment, said cap body 6 is made of an elastic fluid-tight material, e.g., rubber. In an embodiment, said cap body 6 forms a fluid seal directly with the tubular container 2, avoiding an interposition of further seals or sealing elements.

[0076] . In an embodiment, said cap body 6 comprises said reversible access device 7. In an embodiment, said reversible access device 7 is a reversibly pierceable portion of said cap body 6, e.g., a pierceable membrane, known in the field of vacuum tubes.

[0077] . In an embodiment, said reversible access device 7 is a pierceable portion of the cap 5, e.g., a membrane made of a fluid-tight elastomeric material, in which the pierceable portion of the cap 5 is configured to self-seal after being pierced so as to keep the collection chamber 4 closed and fluidly isolated from the external environment.

[0078] . In an embodiment, the at least one reservoir 8, 8’ is fluid-tightly, elastically retained by the cap body 6 in the reservoir housing 15, in both the resting position and the advanced position.

[0079] . In an embodiment, said cap body 6 comprises the at least one separation septum 13, in which the at least one separation septum 13 is a membrane being irreversibly pierceable by the advancement of the reservoir 8, 8’ into the reservoir housing 15.

[0080] . In an embodiment, the cap body 6 is made in one piece with the separation septum 13 and the reversible access device 7, and is shaped to form a seal with the inner walls of the tubular container 2 of the multifunctional device 1 and delimits a cavity with its walls which form the reservoir housing 15. In an embodiment, the cap body 6 is connected by interference to the reservoir 8, 8’ closing the open reservoir end 11 .

[0081] . In an embodiment, each reservoir 8, 8’ protrudes at least partially from the cap body 6 in the direction opposite to the collection chamber 4 and / or the separation septum 13, thereby the reservoir can be easily handled by a user.

[0082] . In an embodiment, said cap 5 comprises a first reservoir 8 and at least a second reservoir 8’ of said at least one reservoir 8, 8’.

[0083] . In an embodiment, the first reservoir 8 is configured to contain a first mixing substance of said at least one mixing substance.

[0084] . In an embodiment, said at least a second reservoir 8’ is configured to contain at least a second mixing substance of said at least one mixing substance.

[0085] . In an embodiment, the second mixing substance is in the liquid state or comprises a gel. In an embodiment, the first mixing substance is in the solid state, for example it is lyophilized. In an embodiment, the second mixing substance is incompatible for storage with the first mixing substance in the same reservoir.

[0086] . In an embodiment, the second mixing substance comprises ingredients in an effective concentration, such as for example: secretome, exosomes, hyaluronic acid, amino acids, gelatin / collagen, hydroxyapatite, tricalcium p-phosphate. Preferably, the exosomes being from any source.

[0087] . For the purposes of the present invention, said at least one mixing substance comprises at least bioactive factors, isolated and purified from colostrum. In accordance with an embodiment, the bioactive factors isolated and purified from colostrum comprise at least growth factors. In accordance with an embodiment, the bioactive factors, isolated and purified from colostrum, comprise at least one of the following biological factors: cytokines, chemotactic factors, stem cell stimulating factors, complement proteins, antibacterial and antiviral factors. In accordance with an embodiment, the bioactive factors, isolated and purified from colostrum, comprise growth factors and other biological factors, e.g., one or more of cytokines, chemotactic factors, stem cell stimulating factors, complementproteins, antibacterial and antiviral factors. In accordance with an embodiment, the bioactive factors, isolated and purified from colostrum are a mixture of bioactive factors isolated and purified from colostrum.

[0088] . In an embodiment, said at least one mixing substance comprises as well as bioactive factors isolated and purified from colostrum, also with other ingredients in effective concentration, such as for example: secretome, exosomes, hyaluronic acid, amino acids, gelatin / collagen, hydroxyapatite, p-tricalcium phosphate, to obtain, in a single closed, sterile and vacuum environment, a (liquid or gel) product with improved bioactive properties for use in treating conditions requiring tissue repair and regeneration, in humans and animals.

[0089] . In an embodiment, said cap 5 comprises said at least one mixing substance contained in said at least one reservoir 8, 8’. In an embodiment, said cap 5 comprises at least bioactive factors, isolated and purified from colostrum, lyophilized.

[0090] . In an embodiment, the multifunctional device 1 is vacuum and / or is a vacuum tube, i.e., in which the collection chamber 4 of the tubular container 2 is depressurized and is at a lower internal pressure than the pressure outside the multifunctional device 1. In an embodiment, the multifunctional device 1 is a tube.

[0091] . In an embodiment, the multifunctional device 1 comprises an anticoagulant 16 arranged in the collection chamber 4 of the tubular container 2, in which the anticoagulant 16 preferably forms a layer inside the collection chamber 4, in which the anticoagulant 16 preferably comprises sodium citrate, in which the anticoagulant 16 is in a predefined amount from 0.5 ml to 2 ml, preferably 1 ml. In an embodiment, the anticoagulant 16 comprises or is sodium citrate.

[0092] . In an embodiment, the multifunctional device 1 comprises a biological fluid composition separator 17 arranged in the collection chamber 4 of the tubular container 2, preferably at the bottom of the tubular container 2, in which preferably the biological fluid composition separator 17 comprises a thixotropic gel, to keep the plasma layers (platelet-rich plasma layers or PRP and platelet-poor plasma layers or PPP) separated from the red blood cells in the centrifuged whole blood.

[0093] . In an embodiment, the cap 5 comprises a cap notch 18, in which the cap notch 18 is configured to be aligned with a device holder notch 30 during a whole blood sampling.

[0094] . In an embodiment, the multifunctional device 1 is configured to prepare autologous platelet concentrates enriched with bioactive factors, isolated and purified from colostrum. The collection chamber 4 is depressurized so as to allow a collection of whole blood by pressure gradient. The at least one reservoir 8, 8’ is in the closed configuration and contains the mixing substance, in which the mixing substance comprises at least growth factors isolated and purified from colostrum. The multifunctional device 1 comprises an anticoagulant 16 arranged in the collection chamber 4 of the tubular container 2. The multifunctional device 1 comprises a biological fluid compositionseparator 17 arranged in the collection chamber 4 at the bottom of the tubular container 2. For example, the anticoagulant 16 preferably forms a layer inside the collection chamber 4 covering the biological fluid composition separator 17.

[0095] . In an embodiment, the multifunctional device 1 is a medical device.

[0096] . In an embodiment, the bioactive factors isolated and purified from colostrum or the mixture of bioactive factors isolated and purified from colostrum, inserted in the reservoir 8, 8’ of the cap of the multifunctional device 1 for preparing enriched autologous platelet concentrates (EAPCs), can be obtained using the method described for example by P. Sacerdote et al. "Biological components in a standardized derivative of bovine colostrum" - Journal of Dairy Science, Volume 96, Issue 3, 1 March 2013, pages 1745 - 1754) or in patent application WO 2022079684A1 .

[0097] . Other methods for preparing bioactive factors can also be employed, as long as they do not involve steps or operations which could damage biological factors, reducing the activity thereof, such as heat sterilization steps which can lead to the loss of large amounts of biological factors in the final product as a result of degradation caused by high temperatures, or operations at low pH values which can result in the denaturation of many proteins, including several biological factors present in colostrum. Accordingly, for the preparation of the bioactive factors and / or the mixture of bioactive factors isolated and purified from colostrum according to the present invention, extraction processes will be used which allow obtaining a mixture of factors in a biologically active form.

[0098] . In an embodiment, the bioactive factors isolated and purified from colostrum which can be inserted into the reservoir 8 are deprived of immunoglobulins and bacterial endotoxins. For the depletion of immunoglobulins, for example, affinity chromatography can be used, while for the depletion of bacterial endotoxins, positive charge filter cartridges can be used which retain the latter, as they have a negative charge.

[0099] . In an embodiment, the reservoir 8, 8’ can contain, together with the bioactive factors isolated and purified from colostrum, other ingredients in an effective concentration and preferably lyophilized or powdered, capable of further enriching the platelet concentrates. This as long as the other ingredients are compatible with the bioactive factors isolated and purified from colostrum and do not damage them in any way: therefore, they must not be oxidizing substances, substances with pH values away from the neutral value beyond half a point, etc. Alternatively, in the case of substances not compatible with the bioactive factors isolated and purified from colostrum or of a different physical state (for example a liquid or a gel), the at least one reservoir 8, 8’ comprises two separate reservoirs but with the same methods of use.

[0100] . In particular, the reservoir s, 8’ can contain, together with bioactive factors isolated and purified from colostrum, one or more of the following components: secretome, exosomes,hyaluronic acid, amino acids, gelatin / collagen, hydroxyapatite, tricalcium p-phosphate. Preferably, the exosomes being from any source.

[0101] . SECRETOME / EXOSOMES. The term secretome is used generically to describe a group of protein molecules which are secreted or released into the extracellular environment by a wide variety of cells, not just stem cells, and these secretory proteins are involved in important biological processes, including cell proliferation, differentiation, cell migration, morphogenesis, angiogenesis, and wound healing. The secretome comprises several bioactive molecules, a significant part of which is represented by growth factors, cytokines and exosomes. Exosomes represent the most extensively studied group among the three major subgroups of extracellular vesicles or EVs (exosomes, microvesicles, and apoptotic vesicles, or ApoEVs). Among these, exosomes are the smallest vesicles, with a variable diameter of 50-150 nm and a cup-shaped morphology when viewed under an electron microscope.

[0102] . Exosomes are secreted by most cell types including immune cells (B cells, T cells), neuronal cells, epithelial cells, endothelial cells, embryonic cells, and mesenchymal stem cells (MSCs). The uptake of exosomes and the contents thereof by the recipient cells is not accidental, but depends on the interactions between the proteins expressed on the surface of the exosomes and the recipient cells.

[0103] . Exosomes possess a specific molecular load which includes membrane proteins, cytosolic and nuclear proteins, extracellular matrix proteins, lipids, nucleic acids (mRNA, miRNA and DNA), growth factors, cytokines and enzymes, underlining the involvement thereof in the regulation, by means of different molecular mechanisms, of various biological functions.

[0104] . Exosomes have been detected in almost all body fluids, under both physiological and pathological conditions, including: urine, blood, serum, breast milk, amniotic fluid, cerebrospinal fluid, saliva, bile, and lymph. Colostrum also contains exosomes as minor but equally important bioactive components. In particular, it has been observed that exosomes derived from bovine colostrum play a crucial role in cell regeneration, above all they seem to have several beneficial effects on UV-induced aging and skin damage because, during treatment, they have the ability to penetrate the stratum corneum and therefore deliver the contents thereof to the dermis.

[0105] . To date, exosomes appear to be involved in immune response, stem cell maintenance and tissue repair.

[0106] . Recent research has also highlighted the potential use of the content of exosomes, in particular growth factors and miRNAs, as a therapeutic treatment of chronic skin ulcers and hypertrophic scars, as well as the possible role of exosomes in modulating the different steps of the wound healing process. However, in order to perform the functions thereof, the exosomes must be present in a functional amount (at least 5 x 109exosomes / mL).

[0107] . There are several main methods for isolating exosomes: (1 ) Ultracentrifugation, (2) size exclusion (e.g., ultrafiltration or chromatography), (3) immunological separation (e.g., antibody-bead capture), and (4) polymer-based precipitation.

[0108] . The advantages in using each method vary based on the different origin of the exosomes (from cell culture supernatant, colostrum or plasma), the degree of purity to be obtained (protein-exosome ratio, with the aim of minimizing the non-exosomal proteins which can be present in biofluids), yield or quality (preservation of the integrity and function of the exosomes).

[0109] . In an embodiment, the exosomes used can be of any origin. In a preferred embodiment, the exosomes used are isolated and concentrated from bovine colostrum.

[0110] . HYALURONIC ACID. Available in various forms (esters, salts, hydrolyzed, cross-linked, liposomal, etc.), among these sodium hyaluronate is preferred, the sodium salt thereof, for the abilities thereof to restore tissue tone and elasticity, as well as to improve and accelerate the re- epithelialization process.

[0111] . AMINO ACIDS. The main amino acids used by fibroblasts for the construction of collagen are Glycine, L-Proline, L-Leucine and L-Lysine. A mixture of these amino acids stimulates the regeneration of collagen.

[0112] . GELATIN / COLLAGEN. Gelatin is a biomaterial which is extracted from collagen, a natural protein present in the bones and skin of animals. Gelatin has been used in food applications since antiquity and in pharmaceutical applications for decades.

[0113] . The body has a high degree of acceptance for gelatin since it can be fully metabolized and is also naturally present in human tissues in the form of collagen. This explains why medical and pharmaceutical scientists are still discovering new uses for gelatin, both with human and veterinary applications, in addition to the several possibilities of use already existing.

[0114] . Gelatin and collagen bring several benefits to (bio)medical applications by virtue of the properties thereof.

[0115] . Gelatin and collagen are natural and biocompatible, they blend easily with other biopolymers;

[0116] . Gelatin and collagen are generally recognized as safe (GRAS) by the Food and Drug Administration (US FDA) and compliant with the US, European and Japanese Pharmacopoeia.

[0117] . Gelatin and collagen are non-immunogenic.

[0118] . Gelatin and collagen can be functionalized to customize the functions thereof.

[0119] . Gelatin is also widely used as a stabilizing agent in vaccines and parenteral formulations in general, by virtue of the high biocompatibility thereof and also helps to provide protection to drugs and various bioactive molecules. To carry out these functions, however, an ultra-pure gelatin is necessary. In fact, high levels of endotoxins, even in medical grade gelatin, can risk causing anundesired immune reaction.

[0120] . HYDROXYAPATITE. Hydroxyapatite is the main inorganic component of bone and makes up 60-70% of the calcified skeleton and 98% of tooth enamel. It is biocompatible, binds quickly to adjacent hard and soft tissues and has a strong osteoconductive capacity. The clinical indications of hydroxyapatite concern the reconstruction of bone tissue and the lining of endo-osseous dental implants to promote osseointegration. Hydroxyapatite is also used as a component of skin fillers.

[0121] . B-TRICALCIUM PHOSPHATE. Beta tricalcium phosphate is a polycrystalline bioceramic, with osteoconductive properties, which in contact with water releases hydroxyapatite crystals. It is used for the resolution of deep intraosseous periodontal defects (periodontal regeneration), for filling post-extraction bone cavities, for bone regeneration, etc. Beta tricalcium phosphate has a progressive resorption, unlike what occurs with hydroxyapatite, with a release of calcium and phosphorus ions which, for example in the case of intraosseous periodontal defects, contribute to the neo-apposition of bone, cement, and periodontal ligament.

[0122] . The present invention also relates to a cap 5 for a tubular container 2 of a multifunctional device 1 . The cap 5 is fluid-tightly connectable to the tubular container 2, closing the open container end 3 so that the tubular container 2 and the cap 5 enclose a collection chamber 4 configured to be depressurized and / or filled with the biological fluid. The cap 5 comprises a cap body 6 fluid-tightly connectable to the tubular container 2, and a reversible access device 7. The reversible access device 7 is fluid-tightly connected to the cap body 6, in which, when the cap 5 is connected to the tubular container 2 of the multifunctional device 1 , the reversible access device 7 is configured to allow a reversible fluid-tight access to the collection chamber 4 without separating the cap 5 from the tubular container 2, to depressurize the collection chamber 4 and / or to collect the biological fluid in the collection chamber 4 and / or to extract the biological fluid from the collection chamber 4. Advantageously, the cap 5 comprises at least one reservoir 8, 8’ fluid-tightly connected to the cap body 6. Each reservoir 8, 8’ delimits a respective containment chamber 9 configured to contain at least one mixing substance. Each reservoir 8, 8’ is in a closed configuration in which the respective containment chamber 9 is closed and fluidly isolated from the external environment, and when the cap 5 is connected to the tubular container 2 of the multifunctional device 1 , from the collection chamber 4. Each reservoir 8, 8’ is configured to be operated and opened on command, passing from the closed configuration to an open configuration, in which when the cap 5 is connected to the tubular container 2 of the multifunctional device 1 , the respective containment chamber 9 is open and in communication, e.g., in fluid communication or at least in fluid communication, with the collection chamber 4, keeping the cap 5 fluid-tightly connected to the tubular container 2. In an embodiment, the cap 5 comprises one or more features previously described in the embodiments of the multifunctional device 1. In an embodiment, the cap 5 comprises three distinct parts: the fluid-tight cap body 6, which encloses thereversible access device 7 and the reservoir housing 15 in a single preformed piece; the at least one reservoir 8, 8’; and a safety cap 31. In an embodiment, the cap 5 consists of said three distinct parts.

[0123] . The present invention further relates to a device holder, or sleeve 19. The device holder 19 is configured to sample biological fluid when connected to a multifunctional device 1 .

[0124] . The device holder 19 comprises a tubular container body 20 configured to accommodate a multifunctional device 1 according to any one of the embodiments described above, in which the at least one reservoir 8, 8’ of the cap 5 of the multifunctional device 1 protrudes at least partially with respect to the reversible access device 7 in the direction opposite to the tubular container 2 of the multifunctional device 1 .

[0125] . The tubular container body 20 is open at a distal end thereof to allow the insertion of the multifunctional device 1 .

[0126] . The tubular container body 20 comprises a bottom wall 21 which closes, at least partially, the tubular container body 20 at a proximal end thereof.

[0127] . The bottom wall 21 is configured to form an end-of-stroke surface 22 against which the cap 5 of the multifunctional device 1 can abut during the insertion of the multifunctional device 1 into the device holder 19.

[0128] . The bottom wall 21 delimits at least one sampling through hole 23, in which the sampling through hole 23 is configured to be aligned with the reversible access device 7 of the multifunctional device 1 .

[0129] . The bottom wall 21 comprises a female connector 24, e.g., of the Luer type, delimiting said sampling through hole 23, in which the female connector 24 and the sampling through hole 23 are configured to be connected to an outlet needle 29 of a sampling tube assembly 26.

[0130] . The bottom wall 21 delimits at least one reservoir through-opening 25 configured to be crossed by the at least one reservoir 8, 8’ avoiding the tubular container body 20 from interfering with the at least one reservoir 8, 8’ during the insertion of the multifunctional device 1 into the device holder 19.

[0131] . The present invention further relates to a kit or kit of parts.

[0132] . The kit comprises a multifunctional device 1 according to any one of the embodiments described above. In an embodiment, the at least one reservoir 8, 8’ of the cap 5 of the multifunctional device 1 protrudes at least partially with respect to the reversible access device 7 in the direction opposite to the tubular container 2 of the multifunctional device 1 .

[0133] . The kit comprises a device holder 19 according to one or more of the embodiments described above.

[0134] . The kit comprises a sampling tube assembly 26 comprising a sampling tube 27, an outlet needle 29 and an inlet needle 28 connected to opposite ends of the sampling tube 27, in whichthe inlet needle 28 is preferably a butterfly needle for sampling whole blood from a patient, in which the outlet needle 29 is configured to pass through the sampling through-hole 23 and connecting to the female connector 24. In an embodiment, the sampling tube assembly 26 is a butterfly needle for venous sampling with pre-assembled Luer adapter, to be inserted into the decentered Luer hole of the device holder 1 , e.g., bell-shaped.

[0135] . The multifunctional device 1 can be inserted into the device holder 19 by aligning the sampling through-hole 23 to the reversible access device 7 of the multifunctional device 1 , and aligning the at least one reservoir 8, 8’ to the at least one reservoir through-opening 25, avoiding the reservoir 8, 8’ from interfering with the bottom wall 21 of the device holder 19 during an insertion of the multifunctional device 1 into the device holder 19.

[0136] . In accordance with an embodiment, the cap 5 comprises a cap notch 18 and in which the device or sleeve holder 19 comprises a sleeve notch 30, in which the cap notch 18 is configured to be aligned with the sleeve notch 30 to allow a correct insertion of the multifunctional device 1 into the device or sleeve holder 19.

[0137] . In an embodiment, the kit comprises a bench-top centrifuge for treating blood, preferably of the Low Speed type with oscillating arms, provided with a rotor, preferably with 4 tilting ampoules of suitable size to accommodate one or more multifunctional devices 1 or tubes.

[0138] . In an embodiment, the kit comprises a counterweight (rocker arm) for the bench-top centrifuge, of shape and weight corresponding to the multifunctional device 1 , to be used during the centrifugation step.

[0139] . In an embodiment, the kit comprises a syringe with a needle of varying dimensions (for subcutaneous, intramuscular, intra-articular use, etc.) for sampling, through the pierceable rubber membrane or reversible access device 7 of the cap 5, APCs enriched during activation, and then administering them again in liquid form at the site to be treated of the patient or, alternatively, placing them in a mold to obtain the enriched platelet clot in the desired shape and dimensions.

[0140] . The present invention further relates to the use of bioactive factors isolated and purified from colostrum as platelet activators. In accordance with an embodiment, the use of bioactive factors isolated and purified from colostrum as platelet activators allows determining platelet degranulation and platelet gel formation. In accordance with an embodiment, the use of bioactive factors isolated and purified from colostrum as platelet activators allows obtaining a platelet clot. In accordance with an embodiment, the use of bioactive factors isolated and purified from colostrum as platelet activators allows determining a platelet clot, avoiding the use of further platelet activators, e.g., platelet activators of known type.

[0141] . Advantageously, it is possible to use the bioactive factors isolated and purified from colostrum both as enriching elements and as activators, allowing the platelet clot to be obtainedwithout necessarily resorting to the use of platelet activators of known type by virtue of the action of the bioactive factors isolated and purified from colostrum.

[0142] . The present invention also relates to a method for preparing enriched autologous platelet concentrates (Platelet-Rich Plasma or PRP or platelet gel, Leukocyte- Rich PRP or L-PRP, Plasma Rich in Growth Factors or PRGF, Platelet-Poor Plasma or PPP, etc.). According to an operating mode, such autologous platelet concentrates are enriched for medical use in the treatment of conditions requiring tissue repair and regeneration, in humans and animals. According to an operating mode, such autologous platelet concentrates are enriched with bioactive factors isolated and purified from colostrum and / or a mixture of bioactive factors isolated and purified from colostrum, preferably in which the bioactive factors comprise at least growth factors. According to an operating mode, such autologous platelet concentrates are enriched, in addition to bioactive factors isolated and purified from colostrum, also with other ingredients in effective concentration, such as for example: secretome, exosomes, hyaluronic acid, amino acids, gelatin / collagen, hydroxyapatite, p-tricalcium phosphate, to obtain, in a single closed, sterile and vacuum environment, a (liquid or gel) product with improved bioactive properties for use in the treatment of conditions requiring tissue repair and regeneration, in humans and animals. Preferably, the exosomes being from any source.

[0143] . The method comprises the step of providing a multifunctional device 1 according to any one of the previously described embodiments, in which the collection chamber 4 is depressurized so as to allow a collection of a predefined amount of whole blood by pressure gradient, in which the multifunctional device 1 comprises an anticoagulant 16 arranged in the collection chamber 4 of the tubular container 2, in which the multifunctional device 1 comprises a biological fluid composition separator 17 arranged in the collection chamber 4 on the bottom of the tubular container 2, in which the anticoagulant 16 preferably forms a layer inside the collection chamber 4 covering the biological fluid composition separator 17, in which the at least one reservoir is in the closed configuration and contains the mixing substance, in which the mixing substance comprises at least bioactive factors isolated and purified from colostrum.

[0144] . The method comprises the step of collecting 1 whole blood in the multifunctional device. In an operating mode, to sample whole blood from the patient, according to procedures known to those skilled in the art, the multifunctional device 1 is used by inserting it into the bell-shaped device holder or sleeve 19, to which the sampling tube assembly 26 has been connected, taking care not to pierce the reversible access device 7, or the rubber membrane, of the cap 5 of the multifunctional device 1 with the inner part of the outlet needle 29 before the tip of the inlet needle 28 has been inserted into the patient’s vein. Preferably, during this operation the notches 18, 30 on both the cap 5 of the multifunctional device 1 and on the device holder 19 are aligned. Then it will be possible to proceed with the venous sampling from the patient according to the techniques known in the art. Inthis step, after inserting the inlet needle 28 into the patient’s vein, the vacuum multifunctional device 1 is pushed inside the device holder 19 until the inner part of the outlet needle 29 has passed through the reversible access device 7, or pierced the rubber membrane, of the cap 5 of the multifunctional device 1 ; preferably during this maneuver the reservoir 8, 8’ of the cap 5 of the multifunctional device 1 is inserted into the special opening 25 of the device holder 19. The vacuum present inside the collection chamber 4 of the multifunctional device 1 is adjusted to automatically aspirate the amount of whole blood necessary to fill it (from 10 mL to 15 mL, preferably 12 mL).

[0145] . In an operating mode, at the end of the venous sampling, the inlet needle 28 is extracted from the patient and the multifunctional device 1 is disconnected from the device holder 19, preferably bell-shaped.

[0146] . In an operating mode, the method comprises the step of inverting the multifunctional device 1 at least three times, preferably 5 times, to mix the whole blood sampled from the patient with the anticoagulant 16, at this point both present in the collection chamber 4 of the multifunctional device 1 . In an embodiment, the anticoagulant 16 comprises at least 1 ml of sodium citrate.

[0147] . In an operating mode, the method comprises a step of centrifuging the multifunctional device 1 , containing the whole blood with the anticoagulant. For example, the method comprises the step of arranging the multifunctional device 1 in a centrifuge for tubes, balancing the weight thereof.

[0148] . In an operating mode, the method comprises the step of centrifuging the multifunctional device 1 , containing the whole blood with the anticoagulant, for at least 5 minutes, preferably less than 15 minutes, preferably 7 minutes, preferably at a speed of at least or more than 3000 rpm, preferably 4000 rpm, to obtain an autologous platelet concentrate (PPP+PRP) comprising platelet-containing plasma separated from red blood cells by the biological fluid composition separator 17, preferably a thixotropic gel. In an operating mode, if it is desired to obtain types of APCs other than PRP, it will be sufficient to remove with a syringe, through the pierceable rubber membrane or reversible access device 7 of the cap 5 of the multifunctional device 1 , a portion of the platelet-poor plasma fraction (PPP) and subject the multifunctional device 1 to further centrifugation (hard spin).

[0149] . The method comprises the step of opening the at least one reservoir 8, 8’ on command putting in communication, e.g., in fluid communication or at least in fluid communication, the containment chamber 9 of the at least one reservoir 8, 8’ and the collection chamber 4 of the tubular container 2 in fluid communication, releasing the mixing substance into the autologous platelet concentrate, without separating the cap 5 from the tubular container 2 of the multifunctional device 1 . In an operating mode, the operator will be able to push the free end of the at least one reservoir 8, 8’ present in the cap 5 of the multifunctional device 1 downwards to drop the lyophilized powder (the bioactive factors isolated and purified from colostrum and possibly other compatible substances) inside the tubular container 2 of the multifunctional device 1 where the APCs themselves are present.In an operating mode, during the step of opening the at least one reservoir 8, 8’ by putting the containment chamber 9 of the at least one reservoir 8, 8’ and the collection chamber 4 of the tubular container 2 in communication, access to the collection chamber 4 through the reversible access device 7 is avoided. In an operating mode, the step of mixing the mixing substance in the autologous platelet concentrate, obtaining an enriched and activated autologous platelet concentrate occurs inside the multifunctional device 1 , e.g., in the collection chamber 4 of the multifunctional device 1 .

[0150] . In an operating mode, the method comprises the step of gently stirring the multifunctional device 1 to mix the mixing substance in the autologous platelet concentrate, obtaining the enriched and at the same time activated autologous platelet concentrate. In an operating mode, the lyophilized powder (the bioactive factors isolated and purified from colostrum and possibly other compatible substances) immediately dissolves, mixing with the APCs and forming the enriched APCs (EAPCs), all in a single closed, sterile and vacuum environment.

[0151] . In an operating mode, the method comprises the step of inserting a needle of a syringe, for example provided with the kit of parts, through the reversible access device 7, for example through the pierceable rubber membrane, of the cap 5 of the multifunctional device 1 , to aspirate the enriched APCs during the activation step. Such a step lasts about 7-10 minutes, during which the operator will have time to administer them again in liquid form in the patient's site to be treated or, alternatively, place them in a previously prepared mold to obtain the enriched platelet clot with the desired shape and size, this will then be placed on the corresponding lesion of the patient.

[0152] . In an operating mode, the activation of the autologous platelet concentrate and the formation of the enriched platelet clot can also be obtained inside the multifunctional device 1 .

[0153] . In an embodiment, the activation of the autologous platelet concentrate and the formation of the enriched platelet clot is obtained without the use of platelet activators of known type by virtue of the action of the bioactive factors isolated and purified from colostrum.

[0154] . In an embodiment, the method comprises the step of collecting the enriched and activated autologous platelet concentrate for cosmetic use.

[0155] . In an embodiment, the method comprises the step of collecting the enriched and activated autologous platelet concentrate for medical use in the treatment of conditions requiring tissue repair and regeneration, in humans and animals.

[0156] . In an operating mode, the at least one mixing substance comprises as well as bioactive factors isolated and purified from colostrum, also for example: secretome, exosomes, hyaluronic acid, amino acids, gelatin / collagen, hydroxyapatite, p-tricalcium phosphate, to obtain, in a single closed, sterile and vacuum environment, a (liquid or gel) product with improved bioactive properties for use in treating conditions requiring tissue repair and regeneration, in humans and animals. Preferably, the exosomes are from any source.

[0157] . By virtue of the suggested solutions, it is possible to solve the technical problems related to blood sampling, the separation thereof into fractions and the integration of the growth factors present in the PRP and PPP with those derived from colostrum, all carried out in a single environment (the multifunctional device 1 ) which is closed, sterile and vacuum, so as to achieve the goals of extreme safety and high efficiency.

[0158] . By virtue of the suggested solutions, it has been surprisingly discovered that the bioactive factors isolated and purified from colostrum also possess the property of activating platelets, as a result, with the described solutions it is possible to obtain the platelet clot without resorting to the use of calcium chloride (typically used in amounts of about 0.2 mL of 10% CaCh, with or without thrombin) for platelet activation.

[0159] . By virtue of the suggested solutions, it is possible to carry out the following steps all in a single closed, sterile and vacuum environment, i.e., inside the multifunctional device 1 : sample whole blood, mix it with an anticoagulant, centrifuge it with a blood composition separator to keep the plasma layers (plasma layers rich in platelets or PRP and plasma layers poor in platelets or PPP) separated from the red blood cells in the centrifuged whole blood, mix isolated and purified bioactive factors from the colostrum (and possibly other substances) exclusively with the PRP / PPP fraction obtained from the separation of the centrifuged whole blood.

[0160] . By virtue of the suggested solutions, it is possible to obtain, in a single closed, sterile and vacuum environment, a product with improved bioactive properties for use in treating conditions which require tissue repair and regeneration in humans and animals.

[0161] . By virtue of the suggested solutions, it is possible to carry out under aseptic conditions: sampling whole blood, separating red blood cells from white blood cells, plasma and platelets (PPP and PRP), mixing the latter fraction with the bioactive factors isolated and purified from colostrum and the simultaneous activation of the enriched PRP thus formed (or of the other APCs).

[0162] . By virtue of the suggested solutions and the use of a completely closed system, the multifunctional device 1 , avoiding the frequent movements of blood / plasma / APCs between syringes and tubes of the prior art and vice versa, it is possible to obtain Enriched Autologous Platelet Concentrates (EAPCs) and activated, which are totally sterile and free of any environmental pollutants.

[0163] . By virtue of the reversible access device 7, e.g., the pierceable rubber membrane, it is possible to insert the distal part of the outlet needle 29 to be used, together with the sleeve 19 and the other parts of the sampling tube assembly 26, to sample whole blood from the patient; to insert the syringe needle to be used to aspirate the unnecessary portion of PPP and thus obtain APCs other than PRP; to insert the syringe needle to be used for sampling enriched APCs from the multifunctional device 1 and for the subsequent administration thereof to the patient or, alternatively, for theplacement thereof in a mold to obtain (after about 7-10 minutes) the enriched platelet clot of the shape and dimensions required.

[0164] . The enriched APCs obtained according to the solutions described above can then be used in the liquid state to be injected at the joint, muscle, subcutaneous level or wherever a regenerative activity is necessary for injured tissues, or for example, they can be used in the form of a sterile spray to treat injuries over large areas of the body. Alternatively, an enriched platelet clot in the desired shape and size can be made to be applied to wounds, sores, ulcers, fistulas, or even to internal lesions to promote healing, such as for example following partial removal of organs or in the source of a fracture.

[0165] . By virtue of the completely closed device, the multifunctional device 1 , the non-use of the normal tubes to prepare enriched APCs and the absence of frequent movements of blood between syringes and tubes and vice versa, the enriched APCs obtained according to the described solutions are totally sterile and free of any environmental pollutants.

[0166] . The invention has been mainly described above with reference to some embodiments. However, as will be readily appreciated by those skilled in the art, other embodiments than those described above are equally possible within the scope of the invention, without however departing from the scope of the claims below.LIST OF REFERENCE NUMERALSMultifunctional device tubular container open container end collection chamber cap cap body reversible access device reservoir second reservoir containment chamber separation device open reservoir end passageway separation septum reservoir piercing portion reservoir housing anticoagulant biological fluid composition separator cap notch Device holder tubular container body bottom wall end-of-stroke surface sampling through-hole female connector reservoir through-opening sampling tube assembly sampling tube inlet needle outlet needle sleeve notch safety cap

Claims

CLAIMS1. A multifunctional device (1 ) for collecting a biological fluid, for example whole blood, comprising:- a tubular container (2) delimiting an open container end (3),- a cap (5), wherein the cap (5) is fluid-tightly connected to the tubular container (2), closing the open container end (3) so that the tubular container (2) and the cap (5) enclose a collection chamber (4) configured to be depressurized and / or filled with the biological fluid, wherein the cap (5) comprises a cap body (6) fluid-tightly connected to the tubular container (2), wherein the cap (5) comprises a reversible access device (7), wherein the reversible access device (7) is fluid-tightly connected to the cap body (6), wherein the reversible access device (7) is configured to allow a reversible fluid-tight access to the collection chamber (4) without separating the cap (5) from the tubular container (2) to depressurize the collection chamber (4) and / or collect the biological fluid in the collection chamber (4) and / or extract the biological fluid from the collection chamber (4), wherein the cap (5) comprises at least one reservoir (8, 8’) fluid-tightly connected to the cap body (6), wherein each reservoir (8, 8’) delimits a respective containment chamber (9) configured to contain at least one mixing substance, wherein each reservoir (8, 8’) is in a closed configuration in which the respective containment chamber (9) is fluidly isolated from the collection chamber (4), wherein each reservoir (8, 8’) is configured to be operated and opened on command, switching from the closed configuration to an open configuration in which the respective containment chamber (9) is open and in communication with the collection chamber (4) keeping the cap (5) fluid-tightly connected to the tubular container (2).

2. A multifunctional device (1) according to the preceding claim, wherein each reservoir (8, 8’) is integral with the cap body (6).

3. A multifunctional device (1) according to any one of the preceding claims, wherein each reservoir (8, 8’) is configured to be operated and opened on command, switching from the closed configuration to the open configuration keeping the cap (5) fluid-tightly connected to the tubular container (2) and avoiding access to the collection chamber (4) through the reversible access device (7).

4. A multifunctional device (1 ) according to any one of the preceding claims, wherein each reservoir (8, 8’) delimits a respective open reservoir end (11 ), wherein the cap (5) comprises at least one separation device (10),wherein said at least one separation device (10), in said closed configuration of said reservoir (8, 8’), is fluid-tightly connected to the reservoir (8, 8’), closing the respective open reservoir end (11) and fluidly isolating the respective containment chamber (9) from the collection chamber (4), and wherein said at least one separation device (10), in said open configuration of said reservoir (8, 8’), is configured to delimit at least one passageway (12), putting the respective containment chamber (9) in communication with the collection chamber (4), wherein said at least one separation device (10) is operable on command, forming said at least one passageway (12) so as to switch on command the respective reservoir (8, 8’) from the closed configuration to the open configuration, keeping the cap (5) fluid-tightly connected to the tubular container (2).

5. A multifunctional device (1 ) according to the preceding claim, wherein the cap body (6) delimits a reservoir housing (15) in which the reservoir (8, 8’) is fluid-tightly housed, wherein the separation device (10) is fluid-tightly connected to the cap body (6), delimiting the reservoir housing (15) in the direction of the collection chamber (4), wherein the reservoir (8, 8’), under an external thrust action, is configured to fluid-tightly advance in the reservoir housing (15) in the direction of the collection chamber (4) from a resting position to at least an advanced position, wherein, in said resting position, the reservoir (8, 8’) is in said closed configuration of said reservoir (8, 8’), wherein, in said at least one advanced position, the reservoir (8, 8’) is in said open configuration of said reservoir (8, 8’) in which the reservoir (8, 8’) operates the separation device (10), forming said at least one passageway (12).

6. A multifunctional device (1 ) according to any one of the preceding claims 3 to 4, wherein said at least one separation device (10) comprises at least one separation septum (13) constrained to the cap body (6), wherein the at least one separation septum (13) is fluid-tightly connected to the reservoir (8, 8’) in said closed reservoir configuration, and wherein the at least one separation septum (13) is pierceable on command, forming said at least one passageway so as to switch on command the respective reservoir (8, 8’) from the closed configuration to the open configuration, keeping the cap (5) fluid-tightly connected to the tubular container (2).

7. A multifunctional device (1 ) according to the preceding claim, wherein each reservoir (8, 8’) comprises a piercing portion (14) at least partially delimiting the respective open reservoir end (11),wherein the piercing portion (14) is configured to pierce the at least one separation septum (13) following said external thrust action on the at least one reservoir (8, 8’) against the at least one separation septum (13) in the direction of the collection chamber (4), keeping the cap (5) fluid-tightly connected to the tubular container (2) and keeping the reservoir (8, 8’) fluid-tightly connected to the cap body (6).

8. A multifunctional device (1 ) according to any one of the preceding claims, wherein said cap body (6) is made of an elastic fluid-tight material, for example rubber, wherein said cap body (6) forms a fluid seal directly with the tubular container (2), avoiding an interposition of further gaskets or sealing elements.

9. A multifunctional device (1 ) according to the preceding claim, wherein said cap body (6) comprises said reversible access device (7), wherein said reversible access device (7) is a reversibly pierceable portion of said cap body (6).

10. A multifunctional device (1) according to any one of claims 7 to 9, wherein the at least one reservoir (8, 8’) is fluid-tightly, elastically retained by the cap body (6) in the reservoir housing (15), in both the resting position and the advanced position, wherein said cap body (6) comprises the at least one separation septum (13), wherein the at least one separation septum (13) is a membrane being irreversibly pierceable by the advancement of the reservoir (8, 8’) into the reservoir housing (15).

11. A multifunctional device (1 ) according to any one of claims 7 to 10, wherein the reversible access device (7) and the cap body (6) are made in a single piece, preferably of fluid-tight elastomeric material, for example pre-molded soft rubber.

12. A multifunctional device (1 ) according to any one of claims 7 to 11 , wherein the at least one separation septum (13) and the cap body (6) are made in a single piece, preferably of fluid-tight elastomeric material, for example pre-molded soft rubber.

13. A multifunctional device (1) according to any one of the preceding claims, wherein said cap (5) comprises a first reservoir (8) and at least a second reservoir (8’) of said at least one reservoir (8, 8’), wherein the first reservoir (8) is configured to contain a first mixing substance of said at least one mixing substance and said at least a second reservoir (8’) is configured to contain at least a second mixing substance of said at least one mixing substance.

14. A multifunctional device (1) according to the preceding claim, wherein the second mixing substance is in the liquid state or comprises a gel,wherein the second mixing substance is incompatible for storage with the first mixing substance in the same reservoir, and / or wherein the second mixing substance comprises one or more of secretome, exosomes, hyaluronic acid, amino acids, gelatin / collagen, hydroxyapatite, p-tricalcium phosphate.

15. A multifunctional device (1) according to any one of the preceding claims, wherein said at least one mixing substance comprises at least bioactive factors isolated and purified from colostrum, and / or wherein said cap (5) comprises said at least one mixing substance contained in said at least one reservoir (8, 8’).

16. A multifunctional device (1) according to the preceding claim, wherein said at least one mixing substance is lyophilized.

17. A multi-functional device (1 ) according to any one of the preceding claims, wherein the multifunctional device (1) is configured to prepare autologous platelet concentrates enriched with bioactive factors isolated and purified from colostrum, wherein the collection chamber (4) is depressurized so as to allow a collection of whole blood by pressure gradient, wherein said multifunctional device (1 ) comprises an anticoagulant (16) arranged in the collection chamber (4) of the tubular container (2), wherein said multifunctional device (1) comprises a biological fluid composition separator (17) arranged in the collection chamber (4) at the bottom of the tubular container (2), wherein the anticoagulant (16) preferably forms a layer inside the collection chamber (4) covering the biological fluid composition separator (17), wherein the at least one reservoir (8, 8’) is in the closed configuration and contains the mixing substance, wherein the mixing substance comprises at least bioactive factors isolated and purified from colostrum.

18. A device holder or sleeve (19) for sampling biological fluid, comprising a tubular container body (20) configured to accommodate a multifunctional device (1) according to any one of claims 1 to 17, wherein the at least one reservoir (8, 8’) of the cap (5) of the multifunctional device (1 ) protrudes at least partially with respect to the reversible access device (7) in the direction opposite to the tubular container (2) of the multifunctional device (1 ),wherein the tubular container body (20) is open at a distal end thereof to allow the insertion of the multifunctional device (1 ), wherein the tubular container body (20) comprises a bottom wall (21) partially closing the tubular container body (20) at a proximal end thereof, wherein the bottom wall (21 ) is configured to form an end-of-stroke surface (22) against which the cap (5) of the multifunctional device (1 ) can abut during the insertion of the multifunctional device (1 ) into the device holder (19), wherein the bottom wall (21 ) delimits at least one sampling through-hole (23), wherein the sampling through-hole (23) is configured to be aligned with the reversible access device (7) of the multifunctional device (1 ), wherein the bottom wall (21 ) comprises a connector (24), for example a female Luer-type connector, delimiting said sampling through-hole (23), wherein the connector (24) and the sampling through-hole (23) are configured to be connected to an outlet needle (29) of a sampling tube assembly (26), wherein the bottom wall (21) delimits at least one reservoir through-opening (25) configured to be partially crossed by the at least one reservoir (8, 8’), avoiding the tubular container body (20) from interfering with the at least one reservoir (8, 8’) during the insertion of the multifunctional device (1 ) into the device holder (19).

19. A kit of parts comprising a multifunctional device (1 ) according to any one of claims 1 to 17, wherein the at least one reservoir (8 8’) of the cap (5) of the multifunctional device (1) protrudes at least partially with respect to the reversible access device (7) in the direction opposite to the tubular container (2) of the multifunctional device (1), and a device holder (19) according to claim 18, and a sampling tube assembly (26) comprising a sampling tube (27), an outlet needle (29), and an inlet needle (28) connected to opposite ends of the sampling tube (27), wherein the inlet needle (28) is preferably a butterfly needle for sampling whole blood from a patient, wherein the outlet needle (29) is configured to pass through the sampling through-hole (23) connecting to the female connector (24), wherein the multifunctional device (1 ) is insertable into the device holder (19) by aligning the sampling through-hole (23) with the reversible access device (7) of the multifunctional device (1), and aligning the at least one reservoir (8, 8’) with the at least one reservoir through-hole (25), avoiding the reservoir (8, 8’) from interfering with the bottom wall (21) of the device holder (19) during an insertion of the multifunctional device (1 ) into the device holder (19).

20. A kit of parts according to the preceding claim,wherein the cap (5) comprises a cap notch (18) and wherein the device holder or sleeve (19) comprises a sleeve notch (30), wherein the cap notch (18) is configured to be aligned with the sleeve notch (30) to allow a correct insertion of the multifunctional device (1) into the device holder or sleeve (19), and / or wherein the kit comprises a bench-top centrifuge for treating blood, preferably of the Low Speed type with oscillating arms, provided with a rotor, preferably with four tilting ampoules of suitable size to accommodate the multifunctional device (1 ), a counterweight (rocker arm) for the bench-top centrifuge, of shape and weight corresponding to the multifunctional device (1 ), to be used during the centrifugation step, and / or wherein the kit comprises a syringe with needle for sampling the fluid contained in the collection chamber (4) through the reversible access device (7) of the cap (5) of the multifunctional device (1).

21. Use of bioactive factors isolated and purified from colostrum as platelet activators.

22. Use of bioactive factors isolated and purified from colostrum as platelet activators according to the preceding claim, to determine platelet degranulation and platelet gel formation.

23. Use of bioactive factors isolated and purified from colostrum as platelet activators according to any one of the preceding claims from 21 to 22, to obtain a platelet clot.

24. A method for preparing enriched autologous platelet concentrates comprising the steps of:- providing a multifunctional device (1) according to any one of claims 1 to 17, wherein the collection chamber (4) is depressurized so as to allow a collection of whole blood by pressure gradient, wherein the multifunctional device (1) comprises an anticoagulant (16) arranged in the collection chamber (4) of the tubular container (2), wherein the multifunctional device (1 ) comprises a biological fluid composition separator (17) arranged in the collection chamber (4) at the bottom of the tubular container (2), wherein the anticoagulant (16) preferably forms a layer inside the collection chamber (4) covering the biological fluid composition separator (17), wherein the at least one reservoir (8, 8’) is in the closed configuration and contains the mixing substance,- collecting whole blood in said multifunctional device (1),- mixing the whole blood with the anticoagulant (16),- subjecting the multifunctional device (1 ) to centrifugation, obtaining an autologous platelet concentrate comprising plasma containing platelets separated from red blood cells by means of the biological fluid composition separator (17), preferably a thixotropic gel,- opening the at least one reservoir (8, 8’) on command, putting the containment chamber (9) and the collection chamber (4) in communication, releasing the mixing substance in the autologous platelet concentrate, without separating the cap (5) from the tubular body (2) of the multifunctional device (1),- mixing the mixing substance in the autologous platelet concentrate, obtaining an enriched, activated autologous platelet concentrate, wherein said mixing substance comprises at least bioactive factors isolated and purified from colostrum.

25. A method according to the preceding claim, wherein during the step of opening the at least one reservoir (8, 8’), putting the containment chamber (9) and the collection chamber (4) in communication, access to the collection chamber (4) by means of the reversible access device (7) is avoided.

26. A method according to any one of the preceding claims 24 to 25, wherein the step of mixing the mixing substance in the autologous platelet concentrate occurs in the collection chamber (4) of the multifunctional device (1 ).

27. A method according to any one of the preceding claims 24 to 26, wherein the step of mixing the mixing substance in the autologous platelet concentrate comprises the step of stirring the multifunctional device (1 ).