Patent Application
20170274052
This invention relates to surgical methods and apparatus in general, and more particularly to novel methods and applications for creating bodily-derived compositions and/or implanting bodily-derived compositions into the body during a medical procedure.
In many situations, it may be necessary or desirable to deploy an implant into the body. In some cases, the implant may be relatively stiff or rigid (e.g., an orthopedic implant). In other cases, the implant may be relatively soft and pliable (e.g., a cosmetic implant).
In some cases, the implant may not integrate well with the adjacent tissue. This can be particularly problematic in cosmetic applications, where implant integration with surrounding tissue can be critical for proper tissue support and proper tissue appearance.
The present invention comprises the provision and use of a novel bodily-derived composition.
In one preferred form of the present invention, there is provided a composite biomaterial comprising at least one blood component and adipose-derived stem cells wherein the at least one blood component is bonded to the adipose-derived stem cells as the at least one blood component coagulates.
In another preferred form of the present invention, there is provided a method for making a composite biomaterial comprising:
extracting at least one blood component from a body;
extracting adipose-derived stem cells from a body;
placing the at least one blood component and adipose-derived stem cells in a vessel; and
bonding the at least one blood component to the adipose-derived stem cells as the at least one blood component coagulates.
In another preferred form of the present invention, there is provided a method for treating a patient, the method comprising:
extracting at least one blood component from a body;
extracting adipose-derived stem cells from a body;
placing the at least one blood component and adipose-derived stem cells in a vessel;
activating at least one blood component so that at least one blood component binds to the adipose-derived stem cells as the at least one blood component coagulates; and deploying the composite biomaterial into a human body.
In another preferred form of the present invention, there is provided apparatus for making a composite biomaterial, the apparatus comprising:
In another preferred form of the present invention, there is provided a method for treating a patient, the method comprising:
extracting at least one blood component from a body;
extracting adipose-derived stem cells from a body;
placing the at least one blood component in a vessel;
clotting the at least one blood component in a vessel;
mixing the adipose-derived stem cells with the at least one clotted blood component to form a composite biomaterial; and deploying the composite biomaterial into a human body.
These and other objects and features of the present invention will be more fully disclosed or rendered obvious by the following detailed description of the preferred embodiments of the invention, which is to be considered together with the accompanying drawings wherein like numbers refer to like parts, and further wherein:
This invention relates to aesthetic, surgical, and injection methods and apparatus in general, and more particularly, to a novel method and apparatus for forming, harvesting, and implanting autologous blood-derived fibrin clot biomaterials, optionally combining these with autologous or allogenic cells, synthetic implants, injectable fillers, supports, and the like, and depositing these into the body of a mammal during a medical procedure. The invention also relates to apparatus that induces fluidic motion within a container to elicit selectively-variable implant matrixes as tissue fillers and cell delivery scaffolding means, and biomaterials.
It is also an object of the present invention to provide systems in the form of kits and apparatus to facilitate novel procedures pertaining to forming completely autologous fibrin adipose composite biomaterials by harvesting platelets from blood, harvesting subcutaneous adipocytes, optionally purifying and or disassociating these, combining the blood with the fat in a container or tubing, and inducing motion upon the liquid in a vessel that will trigger clotting by fluidic interaction with the interior of a container.
The creation of a hematoma or fibrin clot is an initial and important phase in wound repair. The fibrin clot is formed from platelets circulating in blood and provides a matrix scaffold as well as a chemotactic stimulus to the various cellular elements involved in wound repair. The fibrin clot is typically a naturally-occurring response to an injury to vascularized tissue.
In the past, several studies have demonstrated the role of platelet rich plasma (PRP) in enhancing tissue healing. PRP has been shown to contain growth factors essential to the natural healing process. However, several shortcomings of PRP have been reported. PRP production typically requires a centrifuge, is expensive, time-consuming, and is inconsistent depending on the patient and the preparation method. In addition, most methods involve a chemical activation step that activates any latent growth factors, including TGF-β1. PRP in liquid form diffuses upon injection into the body, prohibiting long term contact with the injured area. Lastly, PRP in gel form lacks a porous architecture and has no counterpart in nature. This PRP form has been associated with poor outcomes compared to negative controls.
The present invention overcomes the shortcomings of PRP by obviating a centrifuge and additives used to prevent and trigger clotting. The present invention can be practiced with minimal equipment and typically within a 10 minute time period.
The aesthetic skin treatment marketplace has adopted centrifuge-derived platelet rich plasma (PRP) forms for a variety of skin wrinkle applications for skin ageing. These are primarily treated by injection, however, the natural scaffolding for filling voids, suturing, or adding bulk with PRP is lacking unless bulking agents such as collagen or cells are added.
Injected PRP has also gained notoriety in aesthetic surgery as filler and to build epidermis. Autologous adipose tissue transfer (or fat transfer) is another popular treatment for facial skin ageing and for aesthetic augmentations such as for breast augmentation.
Examination of the morphology of the fibrin biomaterial was performed and assessments made as to the presence of the desired growth factors. Using rabbit polyclonal antibodies from abcam, immunohistochemistry was performed to assess the presence of growth factors PDGF-ββ, TGF-β1, VEGF, and FGF basic. For each growth factor, positive staining was compared to a negative control.
The results of this study show that growth factors essential to tissue healing are present in a human blood clot produced by the present invention. This indicates that these man-made blood clots will enhance tissue healing at least as well as PRP. The study also indicates that for all growth factors there is a greater staining intensity around the periphery of the blood clot (where the blood made physical contact with the Hula cup) as compared to the center of the clot. In addition, there is a higher intensity of TGF-β and VEGF staining in the clots than PDGF-ββ and FGF-2. The high presence of VEGF is notable as it is associated with vascular ingrowth which is essential for engrafted cell survivability and matrix regeneration. Conversely, the relatively low presence FGF-2 is associated with minimal production of less desirable fibrosis and scar tissue formation.
Positive staining for VEGF, TGF-β1, and PDGF-ββ indicates that clots produced by the present invention have viable tissue-healing properties. The fact that these bioactive components are present shows that these fibrin biomaterials are capable of enhancing healing at least as well as traditional centrifuge derived Platelet Rich Plasma, without the expensive, time-consuming, and inconsistent preparation process associated with PRP. The present invention produces biomaterials that have similar growth factor content to PRP, and are 3D scaffolds that can be made economically, consistently, and in typically less than 15 minutes. The biomaterial of the present invention also possesses potentially superior tissue ingrowth properties due to its open architecture and slow release of growth factors.
The present invention produces an autologous fibrin clot scaffold that is an ideal biomaterial for tissue regeneration and addresses the deficiencies of the art as an autologous fibrin clot can now be formed into virtually any shape, density and a range of fiber sizes and orientations to enable a variety of target types, payloads and implantation methods. Autologous fibrin biomaterial can provide an optimized structural microenvironment for stem cell therapy that is also laden with a physiologically-balanced mixture of growth factors having elution kinetics refined by millions of years of evolution.
Transplantation of adipose tissue-derived stem cells is a treatment in which stem cells are extracted from adipose tissue, concentrated, and re-injected together with injectable fat. Some cosmetic surgery clinics claiming “breast augmentation by transplantation of adipose tissue-derived stem cells” perform breast augmentation together with injection of drugs for fat retention and prevention of implant necrosis. Conventional breast augmentation procedures include fat injection, hyaluronic acid (Sub-Q) injection, and insertion of a bag prosthesis (artificial bag). Breast augmentation by transplantation of adipose tissue-derived stem cells is a well-accepted technique, however, issues with cell survival and structural integration limit its effectiveness. In an embodiment of the present invention, stem cells collected from the patient's own adipose tissues are concentrated and separated from non-viable lipids and cells, and are added to the fibrin biomaterial, and re-injected into the body together with the fat as a natural composite biomaterial. The fibrin biomaterial provides a cohesive physical biodegradable structural matrix that can be molded into implant shapes or, alternately, can be easily injected into target tissues. Fibrin clot is rich in growth factors, and the presence of vascular endothelial growth factors is significant because an adequate blood supply improves tissue graft viability. Because transplanted stem cells differentiate into new adipose tissue or vascular endothelial cells, this procedure has the advantages of a much higher fat graft survival (retention) rate and longer survival compared with common fat injection. Because fibrin-adipose biomaterials can be made from the patient's own blood and adipose tissue without additives, this represents a safer more cost effective treatment with no concern for rejection or drug side effects.
It is also an object of the present invention to trigger and complete a clotting cascade in vitro, using the patient's own blood and stem cells in order to form a fibrin biomaterial that is directly bonded to stem cells to facilitate both implantation and long-term viability of the implanted cells.
A body of evidence is emerging which points to the role of the microenvironment in influencing stem cell fate. For example, Yin reported: “The expression of tendon-specific genes was significantly higher in hTSPCs growing on aligned nanofibers than those on randomly-oriented nanofibers in both normal and osteogenic media.” PRP and previously known autologous biomaterials have not been repeatably formed in vitro with distinct fiber orientations.
Thus there is a need for a new and improved method and apparatus for forming and harvesting fibrin-adipose biomaterials and for the implantation thereof.
The present invention provides a new and improved method and apparatus for harvesting a fibrin clot and creating and implanting fibrin-adipose biomaterial into the body during a clinical or surgical procedure.
The invention comprises a method and apparatus for formation of a diverse range of biomaterials and extracellular matrix (ECM) from whole blood and cells without additives, centrifugation or filtration. A unique aspect of the invention described herein is that it enables the user to selectively create fibrin biomaterials possessing a diverse range of properties that can be more conducive to repair and regeneration. Specifically, the fibrin scaffolds promote tissue regeneration and repair, can be made with specific microenvironments (fibril size, orientation and spacing), and release concentrated growth factors over a longer period of time than chemically-activated PRP. The present invention comprises a (preferably) closed container to hold blood and fat, application of motion to the container, and a means to trigger the clotting cascade by turbulence, mixing and/or contact with air (such as with roughened glass). It will be evident to those skilled in the art that impellers, pumps or alternate means to induce fluid motion within a container can be utilized to practice the present invention. The present invention enables the user to optimize the fiber orientation of the biomaterial in strips, laminates, and tendon-like structures that match the graft or repair.
In one preferred form of the invention, there is provided apparatus for extracting fibrin from blood so as to form fibrin biomaterials, the apparatus comprising:
a vessel for holding drawn blood;
a lid for selectively closing off the vessel; and
a precipitator connected to the vessel for engaging the drawn blood contained within the vessel and acting as a focal point for the precipitation of fibrin clot.
In another aspect of the invention, novel fibrin biomaterials having fibers are organized into largely parallel coherent strands or strings by forcing them through small openings.
In another aspect of the invention, fibrin is combined with cells or fillers and implanted into the body of a mammal.
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For purposes of the present description, the contents of vessel 60 will hereinafter be referred to as drawn blood, however, it should be appreciated that the present invention is not intended to be limited to only drawn blood, but may also refer to drawn blood mixed with various additives, e.g., adipose-derived stem cells, collagen fibers, etc.
In one embodiment of the present invention, and looking now at
In one preferred embodiment of the invention, vessel 60 is agitated by tilting the vessel from side to side or by rotating the vessel about a center axis 110, or both (e.g., in the manner shown in
In another preferred embodiment of the present invention, and looking now at
If desired, rod 105 may comprise line markings 120 (See
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More particularly, it has been discovered that the characteristics (e.g., shape, consistency, etc.) of the fibrin clot can be significantly influenced by the relative position of rod 105 vis-à-vis the bottom surface 63, or another internal surface, of the vessel.
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On the other hand, where distal end 108 of rod 105 is disposed so that it is spaced from the bottom surface of vessel 63, e.g., in the manner shown in
Where rod 105 is disposed so that its distal tip 108 is located just below the surface 118 of drawn blood 92, e.g., in the manner shown in
Preferably, rod 105 is formed out of a sintered glass rod. Rod 105 may also be formed of another material which is configured to precipitate fibrin clot, e.g., a metal member such as steel, or a non-glass material, or an open-ended tube that traps air, etc.
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Micro bubbles of air play a role in clotting. Air trapped in the submerged frosted or sintered glass 145 forms bubbles on the surface of the glass. These bubbles interact with fibrinogen to promote the contact activation (intrinsic) pathway, initiated by activation of the “contact factors” of plasma by bubble surface tension effects and turbulence. The tissue factor (extrinsic) pathway is later initiated by release of tissue factor (a specific cellular lipoprotein). This cascade enables coherent gels to propagate through drawn blood 92 in the container.
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An alternate method of practicing the present invention comprises the steps of initiating the clotting cascade in vessel 60 shown in
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In another preferred form of the invention, and looking now at
During or after blood is drawn into syringe 405 in the presence of air, a syringe nozzle 425 can be capped, or otherwise sealed, and the slidable precipitator tube 420 can be extended to contact the blood and initiate clotting to create a fibrin mass 422. The vent assembly 430 can comprise another syringe container or drain forming a fluid path communicating between the syringe interior and exterior and can be used by opening the vent and compressing syringe plunger 415 to vent off air and/or remove platelet poor plasma. In the case of fat processing, vent assembly 430 can be used to remove unwanted oil, lipids, and blood, or to enable exchange of air/fluid, bubble formation, suction/aspiration of clot, or addition of flowable materials, drugs, agents.
Alternate ways to use the device could involve separating fractions of blood or fat from either end of the inside of the syringe vessel that have been separated in the syringe vessel by centrifuge. For example, after a clot is formed (in contact with the precipitator), the vent in the precipitator, or in another portion of the plunger assembly can be opened, and the plunger can be advanced forcing platelet poor plasma red blood cells (RBCs) out of the syringe body and holding the clot in the main collection container (which may optionally also comprise a red blood cell filter).
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For example, the present invention can be used to lay down fibrin scaffold fibrils of selected orientations, such as parallel, concentric, or alternating layers as a means to form a breast implant that will resist sagging. Alternating layers can be formed by altering the direction of blood flow over the precipitation means. This can be achieved by subjecting the container to a sequence of varied motions to change the deposition induced alignment of fibril layers.
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After approximately 7 minutes of motion, a clot having multi-oriented fibrin will fill the gap between the bottom of precipitator 100 and the bottom surface 63 of vessel 60, while radially-oriented fibrin will comprise the portion of the clot formed around precipitator 100 as shown in
The biomaterial of the present invention is unique because it enables the user to selectively create a diverse range of natural microenvironment structures, such as solid gels, flowable pastes, and dense fibrous forms having a particularly oriented or amorphous fiber structure. These biomaterials also provide unique benefits, e.g.: 1) the solid or gel fibrin maintains its architecture during implantation; 2) the matrix binds and delivers growth factors naturally over time; 3) fibrin biodegrades over a period of weeks and is replaced with derivative new tissue architecture; 4) the biomaterial allows seeded cells to persist with lower losses due to cell death; 5) the porosity allows motile phenotype healer cells to migrate into the open matrix; 6) the biomaterial is a regenerative mimic, pseudo-blastema, similar to the repair modality of erodeles and salamanders, where fibrin clots provide a scaffold for stem cells to regenerate missing or damaged tissue while minimizing or eliminating fibrosis; and 7) red blood cells, which could cause inflammatory response at the repair, adhere to the present biomaterial.
The present invention represents a significant and novel advance in biomaterials. Concentrated stem cells from marrow, fat, or culture methods can be incorporated into the various fibrin scaffold types by first positioning them in the container with non-anticoagulated blood or marrow, then inducing fluid motion within the device to activate the natural clotting cascade. In clinical practice, there is an attempt to mimic the target tissue fiber size and orientation of these fibrin scaffolds and augmentation materials, where possible.
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In one aspect of the invention, blood is dispensed into the container and optionally agitated for up to 4 minutes to initiate the clotting cascade. Blood is allowed to sit with the glass rod or alternate precipitation means (such as air bubbles) touching the blood. The blood will gel and mold itself to the inside of the container taking on the shape of the mold. A small amount of platelet poor plasma will be expelled, only slightly reducing the volume of the clot gel from the original blood volume. The gel clot can also be formed upon various biomaterials, forming useful composites.
Utilizing the same apparatus described above, when agitation is minimized or ceased after a short period of stirring or agitation and the contents of the vessel 60 or syringe 405 are allowed to remain largely still, approximately 95-100% of the blood will form into a gel that assumes the shape of the container. This gel is coherent enough to be removed and handled, can be applied in sheets, blocks or segments, and is strong enough to be sutured into place. This gel looks and feels like liver and can be cut into useful shapes.
This coherent gel can be molded into varied shapes as it assumes the shape of the container that it forms in. For example, breast implants can be molded to appropriate shapes comprising aliquots of adipose cells. These gel clot constructs can also be broken up, shredded, pulled through a sieve or lumen (such as syringe 10 or blunt tip needle opening 50 shown in
The gel will extravisate platelet poor plasma in handling, or when subjected to manual or physiologic point-load pressure, leaving denser fibrin and concomitant growth factors in-situ. The liquid platelet poor plasma trapped in the gels and expelled into the tissue with the implant is also rich in growth factors and serves as a biocompatible liquid delivery medium.
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An alternative use for this gel clot is to form it in a container directly onto a soft tissue reinforcing implant where growth factor release and intimate fibril formation on the implant surface is desired to promote integration of bone or soft tissue.
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Further, a method and apparatus of forming implants where liquid blood is subjected to motion and mixing to initiate a clotting cascade, and immediately applied to a repair site (such as shown in
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The present invention offers a significant advance in the art of cosmetic and plastic surgery by providing a means to entrap adipocytes in a fibrin delivery material that can be formed into semi-solid gels or shapes, or subsequently broken up to be injected as a stranded liquid bearing fibrin fibrils mixed with fat cells.
In one aspect of the present invention, there is provided an apparatus for extracting fibrin from blood so as to form a fibrin biomaterial, the apparatus comprising:
a vessel for holding drawn blood and additives such as cells;
a lid, cap, plunger or closure means for selectively sealing the vessel;
a precipitator connected to the lid for engaging the drawn blood contained within the vessel and acting as a focal point for the precipitation of fibrin clot; and
a means for imparting fluid motion relative to the container.
In one aspect, the precipitator comprises a rod.
In one aspect, the rod is movably connected to the lid.
In one aspect, the rod is rotatable relative to the lid.
In one aspect, the rod is radially slidable along to the lid.
In one aspect, the rod is movable longitudinally relative to the lid.
In one aspect, the lid is rotatable relative to the vessel.
In one aspect, the distal end of the rod engages the bottom surface of the vessel.
In one aspect, the distal end of the rod is spaced a small distance from the bottom surface of the vessel.
In one aspect, the distal end of the rod is disposed just below the top surface of the drawn blood contained within the vessel.
In one aspect, the rod comprises line markings.
In one aspect, the vessel is transparent and comprises volume markings.
In one aspect, the rod comprises sintered glass that forms and holds air bubbles when immersed in liquid.
In one aspect, the vessel is a syringe for drawing blood, the lid is a plunger movable along the syringe, and the precipitator is a rod along which the plunger moves.
In one aspect, the vessel comprises a mold for shaping a fibrin biomaterial implant.
In one aspect, the means for imparting fluid motion relative to the container comprises a motor and a receiving means for the vessel.
In one aspect, the fluidic motion generator comprises a timer and alert means for selection of a cycle of motion more than 30 seconds, and less than 6 minutes followed by a period of sedation of at least 4 minutes for a cohesive gel biomaterial to form.
In one aspect, the fluidic motion generator comprises a timer for selection of a 6-15 minute cycle of motion for a cohesive gel biomaterial having circumferential fiber orientation to form.
In another aspect of the present invention, there is provided a method for forming fibrin biomaterial, the method comprising:
placing drawn blood and fillers such as live cells in a vessel;
mounting a precipitator to the vessel so that the precipitator extends into the material contained in the vessel; and
agitating the drawn materials so as to cause fibrin clot to form that is directly bonded to the filler materials.
In yet another aspect, the fibrin biomaterial is placed into a vessel or tubing and extruded through a small opening so as to entwine and form parallel-oriented fibrin fibrils into a yarn or string like material into a body cavity.
In yet another aspect, the fibrin biomaterial is extruded through successively smaller openings so it can be dispensed through a small applicator without clogging it.
In yet another aspect of the present invention, there is provided an apparatus for extracting fibrin from blood so as to form a fibrin clot optionally combined with living cells, the apparatus comprising:
a vessel for holding drawn blood and cells;
a lid for selectively closing off the vessel;
a precipitator connected to the lid for engaging the drawn blood contained within the vessel and acting as a focal point for the precipitation of fibrin clot; and
a means to manually or mechanically inducing fluidic motion within a container to elicit selectively variable matrixes as tissue fillers and cell delivery implants and biomaterials.
It will be understood that many changes in the details, materials, steps and arrangements of parts, which have been herein described and illustrated in order to explain the nature of the invention, may be made by those skilled in the art without departing from the principles and scope of the present invention.
This patent application claims benefit of pending prior U.S. Provisional Patent Application Ser. No. 61/923,035, filed Jan. 2, 2014 by Javin C. Pierce for METHOD AND APPARATUS FOR HARVESTING CREATION AND IMPLANTATION OF A FIBRIN CLOT BIOMATERIAL (Attorney's Docket No. PIERCE-16 PROV), which patent application is hereby incorporated herein by reference.
| Number | Date | Country | |
|---|---|---|---|
| 61923035 | Jan 2014 | US |
| Number | Date | Country | |
|---|---|---|---|
| Parent | 14160093 | Jan 2014 | US |
| Child | 15452301 | US |
