DEVICE AND METHOD FOR THE TREATMENT OF AN ARTIFICIAL BONE IMPLANT WITH BLOOD

Abstract

A device for treating an artificial bone implant with blood, the device comprising: a container configured to accommodate an artificial bone implant and to be filled with blood, the container comprising an opening; and a cover configured to cover the opening of the container. Five additional embodiments of the device and methods for using it are described herein.

Description

DEVICE AND METHOD FOR THE TREATMENT OF A BONE IMPLANT ARTIFICIAL WITH BLOOD. CROSS REFERENCES TO RELATED APPLICATIONS. This application claims priority over U.S. provisional patent application no. 62 / 279,480, filed on November 3, 2015. FIELD The present invention relates to artificial bone implants. More particularly, the present invention relates to the treatment of artificial bone implants prior to their placement in a patient's body. BACKGROUND Sometimes it is necessary to treat artificial bone implants, such as dental implants and bone substitutes, with various preparations and substances to improve the rehabilitation of the tissue where the implant is placed. Of particular interest is the treatment of artificial bone implants with preparations that promote accelerated osseointegration. Artificial bone implants are manufactured from biocompatible materials. For example, dental implants are made of titanium. One of the processes that promotes the proper establishment of the artificial bone implant in bone tissue is osseointegration, also known as bone integration. Osseointegration is a direct structural and functional connection between living bone and the surface of an artificial bone implant. In other words, osseointegration can be defined as the formation of a direct interface between an artificial bone implant and bone, without the involvement of soft tissue. This is achieved through structural coupling at the point of contact between the bone and the surface of the artificial bone implant. Bone integration implants have been used to treat edentulism and for head and neck reconstruction, to facilitate the retention of mandibular, maxillary, nasal and orbital auricular implants, and for bone anchored hearing aids. With specific reference to artificial dental implants, osseointegration is the primary requirement for the stability of the installed dental implant. Similar to a traumatic injury to bone tissue, the drilling of an implant cavity leads to distinct phases comprising 1 A cascade of complex physiological mechanisms, similar to the direct healing of fractures, occurs. First, fibrin polymerization and blood clot formation take place due to plasma and cellular hemostasis mechanisms. The blood clot serves as an extracellular support matrix that facilitates the invasion of bone-forming cells and neoangiogenesis. Subsequently, osteogenic cells generate new bone tissue within the edges of the drilled hole, on the surface of the installed implant. Osteoblasts migrate to the surface of the implant cavity, differentiating and leading to the formation of new bone tissue in an appositional manner. The degree of new bone formation at the implant drill hole interface largely dictates the stability of the installed dental implants.After a remodeling phase of three to six months, the surface of the dental implant is covered by 60-70% newly formed bone, which closely reflects the degree of bone integration. It is clear that accelerating the osseointegration process of artificial bone implants is important, for example, to shorten the recovery period after placing an artificial bone implant in bone tissue. Accelerating osseointegration of dental implants is crucial because additional steps are required after implant placement, such as attaching a dental prosthesis—for example, a crown, bridge, or denture—to the implant, or placing an abutment to support a dental prosthesis. However, progressing to these additional steps requires the healing of the tissue surrounding the implant. Accelerated osseointegration of the dental implant shortens the healing time after implant placement and speeds up the entire dental implantation process. One way to accelerate osseointegration of an artificial bone implant, such as a dental implant, is to coat its surface with substances or preparations that promote osseointegration, such as growth factors. Additionally, it is advantageous to treat the artificial bone implant with other types of materials or preparations. Examples of such additional materials or preparations include, but are not limited to, those that affect the healing of tissue surrounding the implant, improve healing time after implantation, and enhance the condition of a patient receiving the implant—for example, antibiotics, analgesics, and similar substances. The devices and methods currently available for the pretreatment of artificial bone implants are cumbersome and time-consuming. Therefore, there is a need for a device and methods for the easy and short pretreatment of an artificial bone implant, for example, a dental implant and bone substitute, after the placement of the artificial bone implant in bone tissue. SUMMARY Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly known to a person skilled in the art to which this invention pertains. Although methods and materials similar to or equivalent to those described herein may be used in the practice or testing of the present invention, suitable materials and methods are described below. In case of conflict, the patent specification, which includes the definitions, governs. Furthermore, the materials, methods, and examples are illustrative only and are not intended to be limiting. According to one aspect of the present invention, a device for treating an artificial bone implant with blood is provided, the device comprising: a container, configured to accommodate an artificial bone implant and to be filled with blood, wherein the container comprises an opening; and a cover, configured to cover the opening of the container. According to one setting, the device is configured to spin. According to another method, the container and cover are configured to maintain a negative air pressure in the container, compared to the ambient air pressure. According to another additional modality, the cover is configured to fit the artificial bone implant so that the artificial bone implant is held by the cover. According to an additional modality, the artificial bone implant is a dental implant. According to another additional embodiment, the container further comprises a separator that separates the container into an upper and a lower part, wherein the separator comprises holes that allow contact of the blood in the upper part with the blood in the lower part. According to an additional modality, the artificial bone implant is a bone material. According to another additional modality, the separator is configured to keep the bone material at the top of the container. According to another aspect of the present invention, a method is provided for coating an artificial bone implant with bone integration accelerators, the method comprising: extracting blood from a patient; transferring the blood to a device for treating a bone implant artificial with blood, wherein the device comprises a container configured to accommodate an artificial bone implant and to be filled with blood, and wherein the container comprises an opening and a cover configured to cover the opening of the container, and wherein the container contains the artificial bone implant; centrifuging the device and removing the implant from the device. According to one modality, the container and cover are configured to maintain a negative air pressure in the container compared to an ambient air pressure, and the blood is transferred to the container due to a negative air pressure in the container. According to another method, after drawing blood from the patient, the blood is centrifuged and the plasma separated from the centrifuged blood is transferred to the device, and where instead of centrifuging the device, the plasma is allowed to clot. According to another additional modality, the plasma is allowed to coagulate in an accelerated manner. According to another additional modality, the coagulation of the blood or plasma is accelerated by agitation or ultrasound, or any combination of these. BRIEF DESCRIPTION OF THE DRAWINGS The methods described herein are presented by way of example only, with reference to the accompanying drawings. With specific reference now to the detailed drawings, it should be emphasized that the particulars shown are illustrative and for the purpose of describing the preferred methods. They are presented to provide what is believed to be the most useful and easily understandable description of the principles and conceptual aspects of the methods. In this regard, no attempt is made to show structural details in more detail than is necessary for a fundamental understanding. The description, taken with the drawings, makes it clear to those skilled in the art how various methods can be implemented. In the drawings: - Figures IA-B illustrate, according to an illustrative modality, a side view and a perspective view, respectively, of a device for treating an artificial bone implant with pure blood. - Figure 2 schematically illustrates, according to an illustrative modality, a system for extracting pure blood from a vein directly from a patient's vein to a device containing an artificial bone implant. Figure 3 schematically illustrates, according to an illustrative embodiment, a preferred embodiment of a method for coating an artificial bone implant with bone integration accelerators prior to implanting the artificial bone implant into a target bone tissue, by means of the present invention. - Figure 4A illustrates a pure blood sample after centrifugation in a test tube in the absence of an anticoagulant agent. - Figure 4B shows a coagulated CGF layer separated from the rest of the centrifuged pure blood. - Figure 4C shows a coagulated CGF layer placed on gauze. Figure 5A illustrates schematically and Figure 6A is a photograph of a device comprising a container covered with a cover to which an artificial bone implant is attached, when the container is filled with pure blood. Figure 5B schematically illustrates and Figure 6B is a photograph of a device comprising a container covered with a cover to which an artificial bone implant is attached, when the container is filled with pure blood after centrifugation. Figure 5C illustrates schematically and Figure 6C is a photograph of an artificial bone implant attached to a device cover after the cover and artificial bone implant are separated from the container, following centrifugation with pure blood. - Figure 7 is a scanning electron micrograph (SEM) of the coating layer covering the artificial bone implant after centrifugation with pure blood, observed in Figure 6C. - Figure 8 is a photograph of another modality of an artificial bone implant coated with CGF after centrifugation with pure blood. - Figure 9 shows graphs of the cumulative release of growth factors from coated implants over time. Figures 10A-D are SEM images of dental implants coated with CGF and cultured with MSCs. Figure 10A) A dental implant. Figure 10B) A dental implant coated with CGF. Figure 10C) A dental implant seeded in MSCs. Figure 10D) A dental implant coated with CGF and incubated with MSCs. Figures 11A-D are SEM images of treated dental implant surfaces. Figure 11A) A dental implant surface. Figure 11B) A dental implant surface coated with CGF. The arrow indicates a platelet. Figure 11C) A dental implant surface seeded with MSCs. The arrow indicates the seeded cell. Figure 11D) A dental implant coated with CGF incubated with MSCs. - Figure 12 is a graph showing the number of MSC growth on implant surfaces for two days. - Figures 13A-C schematically illustrate another illustrative modality of the device configured to cover a dental implant. - Figures 14A-B illustrate a Boyden chamber for evaluating the effect of CGF-coated implants on MSC migration and growth rate. - Figure 15 schematically illustrates another illustrative modality of the device configured to form a putty bone from a bone material, for example, a bone substitute. - Figure 16 schematically illustrates, according to an illustrative modality, a disassembled device that is configured to form a putty bone from bone material, - Figure 17 schematically illustrates, according to an illustrative modality, an assembled device that is configured to form a putty bone from bone material. - Figure 18 schematically illustrates, according to an illustrative modality, a system for extracting pure blood from a vein, directly from a patient's vein into a device that is configured to form a putty bone from bone material. - Figures 19A-I are SEM images of a putty bone 700 prepared for use with device 1 which is configured to form a putty bone from bone material. DESCRIPTION OF PREFERRED MODALITIES Before explaining at least one embodiment of the invention in detail, it should be understood that the invention is not limited in its application to the construction details and arrangement of components set forth in the following description or illustrated in the drawings. The invention may have other embodiments or may be practiced or carried out in various ways. Furthermore, it should be understood that the phraseology and terminology employed herein is for the purpose of description and should not be considered limiting. In the description of the various figures described herein, similar numbers refer to similar parts. The drawings are generally not to scale. For clarity, non-essential elements were omitted from some of the drawings. The term "artificial bone implant," as described herein, refers to any type of artificial bone implant known to the art. Examples of an artificial bone implant include, but are not limited to, an artificial bone implant made of biocompatible material, such as titanium, used in various procedures—for example, an artificial dental implant. Other examples include, but are not limited to, bone substitutes in any form known to the art - powders, granules and the like. One of the currently available methods for accelerating osseointegration of an artificial bone implant is to coat the implant with pure blood before its placement in the target bone tissue. The pure blood contains components, hereafter referred to as "bone integration accelerators," that accelerate osseointegration, such as stem cells, growth factors contained in blood plasma, and similar substances. Preferably, the blood sample used for the treatment of the artificial bone implant is an autologous blood sample, specifically a pure blood sample taken from the patient undergoing the implantation procedure. It should be noted that the term "patient", as described herein, refers to any organism that undergoes an artificial bone implant, specifically any animal patient, including a human patient. The current practice is to immerse the artificial bone implant in a pure blood sample, preferably an autologous pure blood sample, just before placing the implant into the target bone tissue. During immersion, the pure blood, which includes bone integration accelerators, adheres to the implant. The implant is then separated from the blood sample and placed into the target bone tissue. However, this method is recognized as inconvenient, messy, and time-consuming, and it can expose the blood-coated implant to non-sterile conditions that may increase the risk of contamination, for example, with pathogenic viruses and bacteria. This can lead to local infection of the tissue surrounding the implant site and even systemic infections requiring lifelong treatment. The present invention provides a device for treating an artificial bone implant with pure blood, in a simple, fast and easy-to-use manner. The present invention further provides a device for the biological activation of an artificial bone implant surface with pure blood, in a simple, fast and easy-to-use manner. The present invention further provides a device for the biological activation of a surface of an artificial bone implant with pure blood, to accelerate the osseous integration of the artificial bone implant, in a simple, fast and easy-to-use manner. More particularly, the present invention provides a device for the biological activation of an artificial bone implant surface with pure blood, to enhance osteoblast migration, adhesion, proliferation, and differentiation, all key to improving bone integration, as well as shortening the implant site rehabilitation period. The present invention further provides methods for treating an artificial bone implant with pure blood, using the device of the invention. The present invention further provides methods for the biological activation of a surface of an artificial bone implant with pure blood, when using the device of the invention. The present invention further provides methods for the biological activation of a surface of an artificial bone implant with pure blood, to accelerate the osseointegration of the artificial bone implant when using the device of the invention. More particularly, the present invention provides methods for the biological activation of an artificial bone implant surface with pure blood, to enhance the migration, adhesion, proliferation, and differentiation of osteoblasts, all key to improving bone integration, as well as shortening the rehabilitation period of the implant site. It should be noted that for the sake of simplicity only, pure blood will occasionally be referred to hereafter in shorthand as "blood". Figures 1A-B illustrate, according to one illustrative embodiment, a side view and a perspective view, respectively, of a device 1 for treating an artificial bone implant with pure blood. According to a preferred embodiment, the device 1 is configured to allow coating an artificial bone implant with bone integration accelerators present in pure blood. According to one embodiment, the device 1 comprises a container 10 having an opening 15 (not shown) and a cover 20 configured to cover the opening 15 of the container 10. The container 10 is configured to accommodate any artificial bone implant 30 known in the art, in any size and structure. According to a preferred embodiment, the artificial bone implant 30 is made of a biocompatible material, for example, titanium.For simplicity only, the artificial bone implant 30 is occasionally referred to hereafter as "implant 30". Figure 1 further illustrates an illustrative implant 30 in the form of a dental implant 30 fitting into container 10. According to a preferred embodiment, cover 20 is configured to engage the implant 30, such that the implant 30 is held in place by cover 20. According to one embodiment, the height of container 10 is similar to the length of implant 30. According to another embodiment, the height of container 10 is greater than the length of implant 30. implant 30, as illustrated in Figures 1A-B. According to another additional embodiment, container 10 is configured to be filled with pure blood in a manner that permits the immersion of an implant 30 in the pure blood when the implant 30 is attached to cover 20 and cover 20 covers container 10. According to one method, pure blood is transferred to container 10 through opening 15, for example, using a syringe or pipette. However, this method of transferring pure blood to container 10 is tedious, time-consuming, and, more importantly, increases the exposure of the pure blood sample and the artificial bone implant 30 to non-sterile conditions. To overcome this problem, according to another method, pure blood is transferred to container 10 while the container 10 is kept sealed by cover 20. This can be achieved, for example, by maintaining negative air pressure inside container 10 and transferring pure blood to container 10 using a device that penetrates the interior of container 10, for example, through cover 20 or through a lower portion 17 of the container (see Figures 1A-B), or through any part of container 10. It should be noted that the term "negative air pressure" will occasionally be referred to hereafter as "vacuum". Consequently, a component, for example, container 10, that has a negative air pressure will occasionally be referred to as a component in "vacuum", for example, "vacuum" container. Therefore, according to one further embodiment, container 10 and cover 20 are configured to maintain negative air pressure in container 10, relative to ambient air pressure. According to another further embodiment, cover 20 is configured to allow the penetration of a needle-like device into the interior of container 10. According to yet another further embodiment, the negative air pressure in container 10 is at a level that permits the entry of a quantity of pure blood into container 10 sufficient to cover an artificial bone implant 30 held by cover 20.These methods allow the transfer of pure blood to container 10 to cover an artificial bone implant 30 that is retained inside container 10 by cover 20, without the need to open cover 20, thus avoiding exposure of the pure blood and the artificial bone implant 30 to non-sterile conditions. According to one method, pure blood is transferred to the sealed container 10, which has negative air pressure inside, from a pure blood source. Examples of a pure blood source include a syringe containing pure blood, a blood bag, or a vein. patient and the like. A preferred modality of the pure blood source is a vein from a patient. According to another preferred modality, the pure blood is transferred from a vein of a patient undergoing implantation of the artificial bone implant 30; specifically, the pure blood sample used to treat the artificial bone implant 30 is an autologous blood sample. According to one embodiment, a method is provided for coating an artificial bone implant 30 with bone integration accelerators prior to implanting the artificial bone implant 30 into a target bone tissue, the method comprising: - providing a device 1 for treating an artificial bone implant with pure blood, the device comprising a container 10 having an opening 15, and a cover 20 configured to cover the opening 15 and hold an artificial bone implant 30; - attach an artificial bone implant 30 to the cover 20; - provide a pure blood sample; - Fill container 10 with the pure blood sample; - cover container 10 with cover 20 holding the artificial bone implant 30 so that at least part of the artificial bone implant 30 is immersed in the pure blood; - incubate the artificial bone implant 30 in pure blood for a period of time that allows the artificial bone implant 30 to be coated with the bone integration accelerators; - remove cover 20 with the artificial bone implant 30 covered with the bone integration accelerators from container 10; and - detach the artificial bone implant covered with the bone integration accelerators from the cover. When using a device 1 that has negative air pressure in the container 10 to coat an artificial bone implant 30 with bone integration accelerators prior to implantation of the artificial bone implant 30 into a target bone tissue, the following modalities apply. According to one embodiment, a method is provided for storing an artificial bone implant 30 in a device 1 for the treatment of an artificial bone implant with pure blood, under negative air pressure conditions, the method comprising: - providing a device 1 for the treatment of an artificial bone implant with pure blood, the device comprising a container 10 having an opening 15 and a cover 20 configured to seal the opening 15, wherein the container 10 and the cover 20 are configured to maintain a negative air pressure in the container 10, the cover 20 being configured to allow the penetration of a needle-like device into an interior of container 10, and the cover 20 is further configured to support an artificial bone implant 30; - attach an artificial bone implant 30 to the cover 20; - sealing the opening 15 of the container 10 with the cover 20 to which an artificial bone implant 30 is attached, so that the artificial bone implant 30 is contained within the container 10; and - create a negative air pressure in container 10. Creating negative air pressure in container 10 is accomplished by any method known in the art, for example, by removing air from container 10 using a needle-like device that penetrates cover 20, while keeping container 10 sealed by cover 20. The method for storing an artificial bone implant 30 in a device 1 under negative air pressure conditions can be carried out, for example, during the manufacture of devices 1 for treating the artificial bone implant with pure blood containing the artificial bone implant 30 under negative air pressure conditions. Such devices 1 are ready for use, making the treatment of an artificial bone implant 30 with pure blood a quick and easy procedure, thus avoiding exposing the artificial bone implant 30 and the pure blood to non-sterile conditions. The use of a device 1 that is prepared by the method mentioned above, can be in accordance with the following illustrative modalities. According to one embodiment, a method is provided for coating an artificial bone implant 30 with bone integration accelerators prior to implanting the artificial bone implant 30 into a target bone tissue, the method comprising: - providing- a device 1 for the treatment of an artificial bone implant with pure blood, comprising a container 10 sealed with a cover 20, and an artificial bone implant 30 coupled to the cover 20 under negative air pressure conditions; - insert into the interior of container 10 a needle-like device that is continuously connected to a source of pure blood; - allow pure blood to fill the inside of container 10 and cover at least part of the artificial bone implant; - incubate the artificial bone implant 30 in pure blood for a period of time that allows the artificial bone implant 30 to be coated with the bone integration accelerators; - remove cover 20 with the artificial bone implant 30 covered with the bone integration accelerators from container 10; and - detach the artificial bone implant covered with the bone integration accelerators from the cover. In one modality, the pure blood source contains the autologous pure blood sample. In another modality, the pure blood source is a syringe containing the pure blood. According to another additional modality, the pure blood source is a blood bag containing the pure blood. According to a preferred modality, the source of pure blood is a vein from a patient. When the source of pure blood is a patient's vein, preferably the vein of a patient undergoing implantation of the artificial bone implant 30, a needle-like device inserted into the patient's vein is continuously connected by a conduit to a needle-like device inserted inside the container 10 through the cover 20. As a result of the negative air pressure inside the container 10, the pure blood is drawn from the patient's vein, through the conduit, into the container 10. Figure 2 schematically illustrates, according to one illustrative embodiment, a system for extracting pure blood from a vein, directly from a patient's vein into a device 1 containing an artificial bone implant 30. According to a preferred embodiment, the pure blood is extracted from a patient's vein, for example, a vein in a patient's hand 50, by venipuncture as known in the art, using, for example, a hypodermic needle 52, configured to be inserted into the vein, continuously coupled to a sheath 54, configured to receive the pure blood. The sheath is provided with a connector 56, configured to connect the sheath and allow the insertion of a tube 58 inside the sheath 54. The tube 58 comprises a first end, configured to be inserted into a sheath 54 through the connector 56, and a second end configured to penetrate the container 10 of the device.This is achieved, for example, by means of a second needle 60 attached to the second end of the tube 58. The second needle 60 is configured to penetrate the container 10 of device 1, and device 1 is configured to allow the penetration of the second needle 60 through it. This is achieved, for example, by using a container comprising, for example, a rubber membrane 66 at its base, specifically on the side of the container 10 opposite the cover 20 to which the artificial bone implant 30 is attached. The membrane. Rubber tubing 66 is configured to allow the penetration of a second needle 60 into the container 10. In this way, during the insertion of a hypodermic needle 52 into the patient's vein, the penetration of the first end of tubing 58 into the sheath 54, which is continuously connected to the hypodermic needle 52, and the penetration of the second needle 60, which is coupled to the second end of tubing 58, into the container 10, provides a direct route through which pure blood is drawn directly from the patient's vein into the container 10 that holds the artificial bone implant 30. Negative air pressure in the container 10 assists the drawing of blood directly from the patient's vein into the container 10. One advantage of this method is that it eliminates the exposure of the pure blood and the artificial bone implant to non-sterile conditions, and allows the artificial bone implant to be coated with pure blood in a simple, quick, and easy way. During experimentation with the aforementioned method for coating an artificial bone implant 30 with bone integration accelerators before implanting the implant into the target bone tissue, it was surprisingly found that using a pure blood sample devoid of an anticoagulant is more beneficial than using a pure blood sample containing anticoagulants, such as heparin citrate. It was found that clotting the blood on the artificial bone implant 30 results in more efficient coverage of the implant with the bone integration accelerators. Therefore, according to another preferred modality, with regard to the method of coating an artificial bone implant 30 with bone integration accelerators prior to implantation of an artificial bone implant 30 in a target bone tissue, the pure blood sample devoid of anticoagulant agents. During experimentation, it was also found that centrifuging device 1 for the treatment of an artificial bone implant 30 with pure blood, while in the container there is an artificial bone implant 30 covered with pure blood devoid of anticoagulant agents, causes a very efficient coverage of the artificial bone implant 30 with the bone integration accelerators. Therefore, according to an additional modality, device 1 for the treatment of an artificial bone implant with pure blood is set to be centrifuged while the container holds an artificial bone implant covered with pure blood. According to another additional modality, in relation to the method for covering an artificial bone implant 30 with bone integration accelerators, before implanting the artificial bone implant 30 In a target bone tissue, centrifugal incubation of device 1 for a period of time that allows the artificial bone implant 30 to be coated with the bone integration accelerators included. According to another additional modality, the centrifugation is in the range of substantially between 2,500-3,500 g in a time interval of substantially between 7-10 minutes. Figure 3 schematically illustrates, according to an illustrative embodiment, a preferred embodiment of a method for coating an artificial bone implant 30 with bone integration accelerators before implanting the artificial bone implant 30 into a target bone tissue, by using the device 1 of the present invention, the method comprising: - draw blood from a patient (112) - preferably the blood is drawn from a vein of a patient in whose body the artificial bone implant will be placed 30; - transferring the blood to a device 1 containing an implant (114) - preferably the device comprises a vacuum container containing an implant and the blood is transferred to a vacuum container to cover the implant; - centrifuge device 1 (116) - according to a preferred embodiment, the container is centrifuged at a rate of substantially 2,500-3,000 g for substantially 7-10 minutes; and - Remove the device implant. Figure 4A illustrates a pure blood sample after centrifugation in a test tube in the absence of an anticoagulant. During centrifugation, the blood separates into three main layers: an upper layer of plasma, also known as platelet-poor plasma (PPP), a lower layer of red blood cells (RBCs), and a solid middle layer, referred to hereafter as the concentrated growth factor (CGF) layer. The solid CGF layer comprises three parts: an upper white portion (WP), consisting of white blood cells (WBCs) and platelets, a lower red portion (RP), and a middle "leukocyte layer" (BC). Because the pure blood is centrifuged in the absence of an anticoagulant, the CGF layer clots. Figure 4B shows a clotted CGF layer separated from the rest of the centrifuged pure blood. Figure 4C shows a coagulated CGF layer placed on gauze. The biological material for coating dental implants is primarily composed of concentrated autologous growth factors (CGFs). It is prepared from pure venous blood collected in sterile tubes without an anticoagulant. After centrifugation, a dense fibrin clot / block, rich in growth factors, is formed. This fibrin clot / block results from a high concentration of fibrinogen, factor XIII, and thrombin. Thrombin XIII, activated by thrombin, cross-links fibrinogen with the fibrin clot, increasing its stability and strength, and protecting it against plasmin-mediated degradation. Essentially, the reinforced fibrin matrix captures multiple growth factors, such as platelet-derived growth factor, transforming growth factor B, vascular endothelial growth factor, and epidermal growth factor. Figure 5A schematically illustrates and Figure 6A is a photograph of a device 1 comprising a container 10 covered with a cover 20 to which an artificial bone implant 30 is attached, when the container 10 is filled with pure blood 500. Figure 5B schematically illustrates, and Figure 6B is a photograph of, a device 1 comprising a container 10 covered with a cover 20 to which an artificial bone implant 30 is attached, when the container 10 is filled with pure blood after centrifugation, for example, in a range of substantially between 2,500 and 3,500 g for substantially between 7 and 10 minutes. As a result of the centrifugation, the pure blood 500 separates into two main layers - a lower layer 510 comprising red blood cells and platelets, and an upper layer 520 comprising plasma. Figure 5C illustrates schematically and Figure 6C is a photograph of an artificial bone implant 30 attached to a cover 20 of the device 1 after the cover 20 and the artificial bone implant 30 are separated from the container 10, after centrifugation with pure blood 500. The artificial bone implant 30 is covered with part of the top layer 520 comprising plasma. The artificial bone implant 30 is held by the cover 20 inside the container 10 such that, after centrifugation, the artificial bone implant 30 is in contact with the top plasma layer, the middle layer of the leukocyte cover, and some of the top of the bottom layer of red blood cells. Therefore, centrifugation separates the artificial bone implant from most of the red blood cells but allows direct contact of the artificial bone implant primarily with the plasma, white blood cells, and platelets of the pure blood sample. As a result of centrifugation, the plasma and the leukocyte coating adhere to the surface of the artificial bone implant. Therefore, the plasma components of the leukocyte coating adhere to the surface of the artificial bone implant, including bone integration accelerators such as growth factors contained in the plasma layer and stem cells contained in the leukocyte coating layer. Figure 7 is a scanning electron micrograph (SEM) of the coating layer covering the artificial bone implant after centrifugation with pure blood, as observed in Figure 6C. The coating layer comprises a fibrin network (without the interlocked red blood cells due to centrifugation). The coating layer of the artificial bone implant is tightly woven and characterized by a dense fibrin texture with thin fibers and pores of approximately 0.1 micrometers. Figure 8 is a photograph of another type of artificial bone implant coated with CGF after centrifugation with pure blood. The fibrin clot coating the artificial bone implant is easily visible. To clinically justify the use of CGF-coated implants, the biological activity of the coating layer was evaluated using an enzyme-linked immunosorbent assay (ELISA). The cumulative release rate of growth factors from the CGF coating layer was studied in vitro by incubating the coated implants in a medium and quantifying the growth factors released into the medium. Specific growth factors that have a biological effect on cell adhesion, proliferation, and osteogenic differentiation—all essential for osseointegration of the implant—were selected for study. These growth factors are primarily released from platelets that are interlocked within the fibrin network.Clot formation begins during centrifugation, where heavy red blood cells (RBCs) sediment first and therefore do not interlock within the fibrin network, while light white blood cells (WBCs) and platelets sediment later, following fibrin clot formation, and thus become entangled within the network. These platelet-rich concentrates within the fibrin clot differentially release growth factors and affect cell differentiation and function.These growth factors include platelet-derived growth factor (PDGF), which enhances cell growth, blood vessel repair and generation, and collagen production; vascular endothelial growth factor (VEGF), which promotes the growth and new generation of vascular cells; tumor necrosis factor alpha (TNF-α), which is involved in systemic inflammation; transforming growth factor beta 1 (TGF-β1), which enhances epithelial cell growth and neogenesis; vascular cells and wound healing; and insulin-like growth factor 1 (IGF-1), which is crucial for cell healing and growth. The kinetics of the cumulative release of growth factors from the coated implants are presented in Figure 9. Figure 9 shows the cumulative release graphs of growth factors from coated implants over time. The coated implants were incubated in a medium at 37°C for varying time intervals (5 h, 1, 3, 6, 7, or 8 days), and the growth factors released into the medium were quantified. The release assay was conducted using three different donors, each incubated with an implant in a separate vacuum container (v1, v2, and v3). The individual and main results are presented: A) PDGF-AB released over time. B) VEGF released over time. C) TNF-α released over time. D) TGF-β1 released over time. E) IGF-1 released over time. All growth factors were present in the CGF coating layer and were released at a slow rate. The release of PDGF-AB and VEGF appeared to increase over the eight-day period, while the release of TNF-α, TGF-β, and IGF-1 appeared to be constant over time. Future studies will measure the release of growth factors in media containing protease inhibitors, and will also account for the release of degraded growth factors. Growth factor release will be evaluated over an extended period, up to 20 days. SEM was used to characterize the surface of CGF-coated dental implants seeded with cells. Mesenchymal stem cells (MSCs) isolated from bone marrow were seeded onto the CGF-coated implants at a density of 100,000 MSCs / ml / dental implant and then cultured for two days. This was done to verify the effect of the CGF coating on cell adhesion and growth. The samples were fixed, and the three-dimensional morphology of the implant coating and cell distribution were visualized (Figures 10A-D and 11AD). Figures 10A-D are SEM images of dental implants coated with CGF and cultured with MSCs. Figure 10A) A dental implant. Figure 10B) A dental implant coated with CGF. Figure 10C) A dental implant seeded in MSCs. Figure 10D) A dental implant coated with CGF and incubated with MSCs. Figures 11AD are SEM images of treated dental implant surfaces. Figure 11A) A dental implant surface. Figure 11IB) A dental implant surface coated with CGF. The arrow indicates a platelet. Figure 11C) A dental implant surface seeded with MSCs. The arrow indicates the seeded cell. Figure 11D) A dental implant coated with CGF incubated with MSCs. The arrows indicate the seeded cells. The bare surface of the titanium dental implant (Figure 11A) differs from the implant surface coated with CGF, with the layer showing fibrin fibers (Figure 1IB). The cells attached 17 to the bare surface of the implant (Figure 110), but were observed in greater numbers on the surfaces coated with CGF (Figure 11D). After demonstrating the three-dimensional structure of the CGF-coated layer and the rate of growth factor release, the biological activity of the CGF coating on bone marrow-derived MSCs was investigated. Since the CGF coating contains the fibrin matrix and growth factors, MSC adhesion and proliferation on the layer were tested using the AlamarBlue metabolic activity assay. MSCs were seeded at a density of 100,000 MSCs / ml / dental implant, and the number of cells that adhered to the implant surfaces and proliferated over two days was evaluated (Figure 12). Figure 12 is a graph showing the number of MSCs growing on implant surfaces over two days. The CGF coating significantly improved MSC adhesion and proliferation compared to control samples. The biocompatibility of the fibrin matrix and the effect of interblocking factors on cell growth are projected to enhance the osteogenic differentiation of MSCs. The effect of the CGF coating on osteogenic genes was investigated using techniques such as fluorescence-activated cell scanning (FACS) and real-time PCR. Figures 13A-C schematically illustrate another embodiment of Device 1 configured to coat a dental implant. Figure 13A illustrates a vacuum container comprising two tubes. Blood is transferred to the inner tube, into which a dental implant is placed. The longer tube protects the inner tube during centrifugation. Figure 13B illustrates a centrifuged vacuum-assembled container with a silicone cap facing downwards. Figure 13C illustrates a silicone cap from which the blood is transferred. Figures 13 AC illustrate the vacuum container in which a dental implant is placed to cover the root surface coating during centrifugation. The appropriate vacuum pressure within the vacuum container is set to extract the precise amount of blood for optimal coating. Accelerated blood coagulation can be achieved by coating the internal channel of the vacuum container with micronized silicon and silicone. As such, the blood drawn into the vacuum container undergoes a complex coagulation cascade that forms long fibrin strands around the implant, eventually resulting in a homogeneous, network-like texture. Taking into consideration the vacuum and centrifugation parameters (e.g., relative centrifugal force (ref), time, speed, container orientation in the centrifuge, etc.), a coating process is achieved that produces a layer of 400–500 µm. micrometers thick fibrin that traps bioactive components such as platelets and WBCs, but deprives RBCs. To ensure the effectiveness of the procedure and the bioactivity of the coated implant, which is designed for immediate implantation following coating, the CGF coating is further characterized. The growth, proliferation, migration, and differentiation potential of MSCs seeded on the CGF coating layer is studied. MSC migration is evaluated using the Boyden chamber assay. Figures 14A-B illustrate a Boyden chamber for evaluating the effect of CGF-coated implants on MSC migration and growth rate. Figure 14A illustrates the effect of growth factors released from CGF-coated implants on MSC migration. Figure 14B illustrates the effect of growth factors released from CGF-coated implants on MSC growth rate. The cells are placed in the cell culture insert, or in the upper chamber separated by a porous membrane from the lower chamber, which contains the CGF-coated implant. The cells and implant are immersed in a shared serum-free medium. The cells are then allowed to migrate from the upper chamber to the underside of the insert for 4 hours under incubation conditions. Cells on the upper surface of the insert membrane are mechanically removed, and the cells migrating to the underside are fixed, held, and counted. This technique allows for the evaluation of the percentage of MSC migration around the release of growth factors from the CGF-coated implant.The effect of these factors on the growth rate of MSCs in the lower chamber, in the presence versus absence of CGF-coated implants in the upper chamber, can be further evaluated and compared over time (Figure 14B). The expression of osteogenic genes in MSCs seeded in the CGF coating layer is determined using techniques such as FACS and real-time PCR. The osteogenic potential of the CGF-coated MSCs is tested in vitro using the alkaline phosphatase activity assay and by assessing cell mineralization. Small, standard-sized implants with appropriate CGF-coated containers are used to test the studied concepts for accelerating osseointegration, followed by placement of the CGF-coated implants in rat tibia bone tissue. Standard-sized implants are also tested in humans, along with the appropriate container. Suitable for coating with CGF. These coated implants are placed in jawbone tissue of a dog to establish clinical protocols for human patients. Following in vitro studies characterizing MSC differentiation in the presence of CGF coating, feasibility studies are conducted in which CGF-coated implants are transplanted into the rat tibia to assess their rate of osseointegration. Nude Wistar rats are anesthetized, and an incision is made on the right proximal anterior surface of the tibia. Care is taken to preserve the periosteal surface. Holes are drilled through the cortex using a 1 mm drill bit, and the implants are inserted. The skin is closed around the implant without non-absorbable sutures, and pain is managed. Two groups of animals are tested and compared: those receiving CGF-coated implants versus those receiving uncoated implants. The CGF coating procedure is performed using human blood.The osseointegration of the implant is assessed at several time points (e.g., 2, 4, 6, and 10 weeks after implant placement). A total of 40 rats are required for the study. At the end of the experiment, the rats are anesthetized; the tibial bone in which the implant was placed is removed, fixed in formalin, and embedded in paraffin for histological analysis or analyzed with an ex vivo micro-CT scanner to assess bone tissue formation and osseointegration of the implant. The rats are euthanized by intracardiac administration of sodium pentobarbital. The osseointegration of CGF-coated implants is evaluated in a canine model. Two groups of four male beagles, each between two years of age and weighing 15–18 kg, are radiographically examined prior to tooth extraction to rule out any pathology. Two mandibular implants are implanted in each dog, with one group receiving CGF-coated implants and the other receiving uncoated implants. The implant coating is performed using autologous blood, collected from each dog to be treated. The dogs undergo surgery under halothane gas anesthesia. Heart rate, temperature, and respiratory rate are monitored during surgery. The edentulism procedure is evaluated radiographically. Venous blood from the dogs is used for the implant coating.Osseointegration and healing of the bone tissue around the implants are assessed radiographically at 1, 3, and 6 months post-implantation. At the 6-month time point, the implants are also evaluated histologically. Histomorphometric analysis is performed to determine the percentage of bone contact length along the implant. Figure 15 schematically illustrates another embodiment of the device 1 configured to form a putty bone from bone material, for example, a bone substitute. Bone material 600, in the form of powder, fragments, and the like, is placed inside the vacuum container 10. The container 10 is provided with a fixed separator 40 that divides the container 10 into two parts, a lower and an upper part. The separator 40 is provided with holes such that when the container 10 is filled with pure blood 500, the blood 500 in the lower part of the container 10 is in complete contact with the blood 500 in the upper part of the container 10. The bone material 600 is in the upper part of the container 10, above the separator 40.After the vacuum in container 10 is replaced with patient blood 500, container 10 is centrifuged under conditions similar to those described earlier. As a result of centrifugation, the blood 500 is separated into a lower phase 510 comprising red blood cells and platelets, mostly concentrated in the lower part of container 10, below the separator 40, and an upper phase 520 comprising plasma in the upper part of container 10, above the separator 40. The plasma again contains the bone integration accelerators, which adhere to the bone material 600 to form a putty bone 700, ready for use. Figure 16 schematically illustrates, according to an illustrative modality, a disassembled device 1 configured to form a putty bone from bone material. The container 10, illustrative cover components 20, and the separator 40 are illustrated. Figure 17 schematically illustrates, according to an illustrative modality, an assembled device 1 configured to form a putty bone from bone material. The container 10, illustrative cover components 20, and the separator 40 are illustrated. Figure 18 schematically illustrates, according to an illustrative modality, a system for extracting pure blood directly from a patient's vein into a device 1 configured to form a putty bone from bone material. The system components are described in Figure 2, and the device 1 configured to form a putty bone from bone material is as described in Figure 15. Figures 19A-I are SEM images of a putty bone 700 prepared for use with the device Ique configured to form a putty bone from bone material. It is apparent that certain features of the invention, which, for clarity, are described in the context of separate embodiments, can also be provided in combination in a single embodiment. Conversely, several features of the invention that are described, for the sake of brevity, In the context of a single modality, they may also be provided separately or in any suitable subcombination. Although the invention has been described along with its specific embodiments, it is evident that many alternatives, modifications, and variations will be apparent to those skilled in the art. Accordingly, it is intended to encompass all such alternatives, modifications, and variations that fall within the spirit and scope of the appended claims.

Claims

CLAIMS 1. A device for treating an artificial bone implant with blood, the device comprising: a container configured to accommodate an artificial bone implant and be filled with blood, wherein the container comprises an opening; and a cover configured to cover the container opening.

2. The device of claim 1, wherein the device is configured to centrifuge.

3. The device of any of claims 1-2, wherein the container and cover are configured to maintain a negative air pressure in the container, compared to the ambient air pressure.

4. The device of any of claims 1-3, wherein the cover is configured to attach to the artificial bone implant so that the artificial bone implant is held by the cover.

5. The device of claim 5, wherein the artificial bone implant is a dental implant.

6. The device of any of claims 1-3, wherein the container further comprises a separator that separates the container into an upper and a lower part, wherein the separator comprises holes that allow contact of the blood in the upper part with the blood in the lower part.

7. The device of claim 6, wherein the artificial bone implant is a bone material.

8. The device of claim 7, wherein the separator is configured to hold the bone material on top of the container.

9. A method for coating an artificial bone implant with bone integration accelerators, the method comprising: draw blood from a patient; transferring blood to a device for treating an artificial bone implant with blood, wherein the device comprises a container configured to accommodate an artificial bone implant and to be filled with blood, and wherein the container comprises an opening; and a cover configured to cover the opening of the container and wherein the container contains the artificial bone implant; centrifuge the device; and remove the device implant.

10. The method of claim 9, wherein the container and cover are configured to maintain a negative air pressure in the container compared to an ambient air pressure, and the blood is transferred to the container due to a negative air pressure in the container.

11. The method of claim 9, wherein, after drawing blood from the patient, the blood is centrifuged and the plasma separated from the centrifuged blood is transferred to the device, and wherein instead of centrifuging the device, the plasma is allowed to clot.

12. The method of claim 11, wherein the plasma is allowed to coagulate in an accelerated manner.

13. The method of any of claims 9-11, wherein the coagulation of the blood or plasma is accelerated by shaking, or ultrasound, or any combination thereof.

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