METHOD AND APPARATUS FOR CELL TREATMENT

专利摘要:
cell treatment system and apparatus. the present invention relates to systems and apparatus for improving the quality and viability of biological material, such as adipose cells, basal cells or other collected cells or biological components, by treating the biological material with membrane stabilizing / repair agents or the like and / or mechanical removal of components, such as impurities and / or excess treatment agents. the present invention also relates to systems and apparatus for transplanting tissue, such as adipose tissue.
公开号:BR112013002859B1
申请号:R112013002859-9
申请日:2011-08-05
公开日:2020-10-06
发明作者:William G. Austen, Jr
申请人:The General Hospital Corporation D/B/A Massachusetts General Hospital；
IPC主号:

专利说明:

RELATED REQUESTS
[001] This claim claims the benefit under 35 U.S.C.§ 119 (e) of the U.S. provisional application, U.S.S.N. 61 / 371,400, filed on August 6, 2010, entitled "SYSTEM AND APPARATUS FOR CELL TREATMENT", the complete content of which is incorporated herein by reference. FIELD OF THE INVENTION
[002] The present invention relates to methods, systems and apparatus for processing biological material such as collected adipose tissue, adipose cells (adipocytes), basal cells or other cells or tissues. BACKGROUND
[003] Adipose tissue that is collected from the body can be reintroduced or transplanted autologously, for example, as a filling material for cosmetic purposes, such as enhancing certain body features. Adipose tissue can also be used for certain reconstruction and / or functional procedures, such as facial reconstruction, breast reconstruction or enlargement, and injection into the vocal cords to improve voice function. Fat tissue can be collected using conventional liposuction procedures. or other techniques. This collected adipose tissue often includes other agents, such as water or other liquids, free lipids, blood cells, etc., in addition to adipose cells (adipocytes).
[004] Removal of impurities and / or fluid from the collected adipose tissue, before reintroducing the adipocytes into a patient's body can be beneficial for the success of the graft. Several procedures and systems have been developed to achieve this separation or "purification" of the fat. Many procedures (including the one used in the Colleman System) involve the use of a centrifuge to separate the different components of the collected adipose tissue and allow them to be separated. Another system, the PUREGRAFT system, uses a squeegee clamping movement on adipose tissue placed in an IV plastic shaker, to obtain some separation of adipose cells from other components of adipose tissue. The "purified" added cells can then be reintroduced into the body using a syringe, cannula or the like.
[005] A mechanical segregation process, such as a centrifugal or squeegee process, can inflict damage to fat cells and reduce their viability, leading to increased apoptosis and cell death. This damage can originate from excessive forces (for example, shear forces) or pressures that can be formed during purification / segregation procedures. Damaged adipocytes, which die after transplantation, tend to be reabsorbed by the body, reducing the efficiency of the transplant procedure.
[006] The total viability of adipocytes can be improved by exposing them to certain classes of polymers, such as Poloxamer P88, a triblock copolymer, and / or antioxidants such as lipoic acid. Therefore, a collection / purification system, which provides for the treatment of the collected adipose tissue using these agents, which can act as membrane stabilizing agents (MSAs), or cell protectors, to improve the viability of adipose cells, may be desirable, for example, for fat collection and autologous fat transplantation procedures.
[007] Therefore, there is a need for a relatively simple, inexpensive, efficient and safe device, configured to offer some segregation of fat cells from other undesirable tissue components, while limiting mechanical damage to the cells. It is also desirable to obtain a system that does not require a centrifuge or other large or expensive piece of equipment and that can be used in an operating room or doctor's office. It is also desirable to obtain a treatment system for collected adipose tissue, which facilitates the treatment of adipose cells with appropriate agents, to improve the viability of the cells to be transplanted. SUMMARY OF THE INVENTION
[008] Modalities of the present invention offer methods, systems and apparatus for processing biological cells or tissues, which have been removed from an individual, which can optionally be transplanted into the same or a different individual. Such processing may include removing impurities, for example, surplus liquids, free lipids, blood cells or other components of biological material that may be undesirable, and / or adding certain agents to biological material, to obtain certain effects (for example, improve the viability of the graft) and, optionally, remove an excess of these agents, before transplanting the biological material in an individual.
[009] For example, improving the viability of adipose cells and / or other cells found in adipose tissue (eg basal cells) for transplantation can be obtained by mixing adipose tissue collected with a membrane and stabilizing agent (MSA) or cell protector , such as Poloxamer P188 or lipoic acid. (See, for example, International PCT Application No. PCT / US2009 / 005727 and U.S. Patent Application Publication No. 2010/0104542, the contents of which are hereby incorporated by reference in their entirety). The apparatus can optionally enable the separation and adipose cells of undesirable components from the collected biological material (for example, excess fluid, blood, free lipids) using a mechanical force, such as a plunger mechanism with a filter element, or using positive or negative pressure. A force limiting arrangement may be provided with the plunger, to limit the amount of force / pressure applied to the fat cells, thereby limiting the amount of mechanical damage that can be induced. Impurities and / or fluids, such as tumescent fluid or free lipid, can also be removed from adipose tissue by providing for a retention matrix (for example, a matrix composed of an absorbent material and / or an adsorbent material) in contact with the adipose tissue .
[0010] According to some aspects of the invention, devices and systems are provided for processing and, optionally, transplanting biological materials, for example, injecting biological materials into a graft site in an individual. The apparatus typically includes a chamber with at least one outlet, and a pressure generating device, configured and arranged to generate positive pressure within the chamber that is at or below a predetermined threshold. Positive pressure, in general, is sufficient to cause biological material, when present in the chamber, to be discharged through the outlet, and the predetermined threshold is, in general, at a pressure above which biological material is relatively viable. low as a tissue graft, after discharge from the outlet at a graft site in the individual.
[0011] In accordance with some aspects of the invention, apparatus and systems described herein offer improved methods for injecting biological materials into graft sites in an individual. Consequently, methods are presently provided for injecting processed biological materials into one or more graft sites in an individual. In some embodiments, the methods involve loading a biological material into the chamber of an apparatus and causing the pressure generating device of the apparatus to generate a positive pressure within the chamber, which is at or below the predetermined threshold, to cause the material biological material is discharged through the outlet. In general, the predetermined threshold is a pressure above which biological material has relatively low viability as a tissue graft after discharge from the outlet at a graft site in an individual. Decreased viability can result from tissue or cell damage caused by relatively high flow rates, pressures, injection rates and / or shear loads imposed on biological material, particularly cells, during processing or injection.
[0012] In other aspects of the invention, systems are provided for processing biological materials. The systems are typically used to process adipose tissue obtained from an individual in one location, typically as a liposuction product, to transfer autologous fat to the individual in a different location. The processing and transfer of fat is typically performed for cosmetic or reconstructive purposes. Systems typically include a chamber with at least one inlet and at least one outlet, a pressure generating device, configured and arranged to transfer biological material to the chamber through at least one inlet and discharge processed biological material out of the chamber through at least one exit. At least one retention matrix is typically present within the chamber. This retention matrix is typically configured and arranged to contact biological material, when present in the chamber, so that a component (e.g., fluid) of the biological material is retained in the retention matrix. The systems may include a variety of other components, in addition to the retention matrix, for the purpose of removing undesirable components from the collected biological material. For example, systems can include filters, antigen binding regions to separate target antigens (for example, cells, soluble proteins, growth factors) and antioxidants to separate free radicals and can also include components to provide additives to materials biological agents, for example, membrane stabilizing agents. In certain embodiments, the system is a modular system, which includes multiple components (for example, filters, end pieces, cannulas, caps, retaining dies, chambers), which can be used as needed. DEFINITIONS
[0013] "Approximately" or "about", when used in reference to a number, generally includes numbers that are within a 5% range in any direction of the number (greater than or less than the number) , unless defined otherwise or where otherwise evident from the context (for example, where that number exceeds 100% of a possible value).
[0014] "Adipocyte" or "adipose cell" refers to a cell that is capable of synthesizing and / or storing fatty acids. The term includes adipocytes with representative properties of cells present within white fat, yellow fat and / or brown fat. Typically, adipocytes store energy in the form of lipids and release energy reserves in response to hormonal stimulation. Morphologically, adipocytes can appear as swollen cells, with dislocated nuclei and a thin cytoplasmic compartment. An adipocyte can express one or more of the following genes: Adiponecin / Acrp30, FATP4, gAdiponectin / gAcrp30, FATP5, Clathrin Heavy Chain 2 / CHC22, Glut4, FABP4 / A-FABP, Leptin / OB, FATP1, PPAR gamma / NR1C3 , FATP2, or Pref-1 / DLK-1 / FA1. Unless otherwise indicated, the term adipocyte includes lipoblasts, mature adipocytes, pre-adipocytes and basal adipogenic cells.
[0015] "Adipose tissue", as used herein, refers to a tissue, which comprises adipocytes or fat. Typically, adipose tissue includes body fat, fat deposits and / or loose connective tissue with adipocytes. Adipose tissue can include white fat, yellow fat, and / or brown fat. The term adipose tissue also includes tissues and components thereof, which are obtained by liposuction product. Adipose tissue may be unprocessed or processed by the methods, apparatus and systems described herein. In addition to adipocytes, adipose tissue can contain multiple types of regenerative cells, including stromal cells, endothelial precursor, basal cells and other precursor cells. The term "fat tissue" can be used interchangeably with "fat tissue".
[0016] "Biocompatible" refers to a material that is substantially non-toxic to cells in the amounts used, and also does not cause or cause a harmful or unfavorable effect on the recipient's body at the site used, for example, an immunological reaction or unacceptable inflammatory.
[0017] "Liposuction product", as used herein, refers to a mixture consisting of adipose tissue and fluid removed from an individual by liposuction. Liposuction product may contain tumescent fluid used during a liposuction procedure. Consequently, the liposuction product may contain lycaine (or another local anesthetic) and / or epinephrine (or another related hormone).
[0018] "Membrane stabilizing agent (MSA)", as used herein, refers to an agent that stabilizes a cell's membrane to prevent injury. MSAs include agents that seal and / or stabilize the membrane of cryopreserved cells, for example, after thawing and, consequently, improve the viability of cryopreserved cells after thawing. MSAs also include agents that prevent injury to adipocytes during adipose tissue processing, for example, during adipose tissue processing for a fat transplant procedure. Typically, MSAs include a nonionic polymer, for example, a nonionic polyether, which interacts with the phospholipid double layer of a cell.
[0019] "Basal cell", as used at present, refers to any cell that, under the right conditions, gives rise to a more differentiated cell. The term "basal cell" includes basal totipotent, pluripotent, multipotent, oligopotent and unipotent basal cells. The term "basal cell" also includes cells (e.g., adult somatic cells), which have been reprogrammed, such as, for example, induced pluripotent basal cells commonly referred to as iPSor iPSCs cells). Non-restrictive examples of basal cells include basal mesenchymal cells, hematopoietic basal cells, stromal cells and basal cells derived from adipose. Basal cells can also be characterized by a capacity to be self-renewing, to regenerate a tissue (for example, adipose tissue), to give rise to other differentiated cells (for example, adipocytes) and / or to produce colony forming units in various laboratory systems.
"Individual", as used herein, refers to an individual to whom an agent is administered, for example, for experimental, diagnostic and / or therapeutic purposes. Preferred individuals are mammals, for example, pigs, mice, rats, dogs, cats, primates or humans. In certain modalities, the individual is a human being. The animal can be a male or female at any stage of development. In certain embodiments, the individual is an experimental animal, such as a mouse or rat. An individual in the care of a doctor or other health care worker can be referred to as the "patient". BRIEF DESCRIPTION OF THE DRAWINGS
[0021] Figure 1 shows a non-restrictive modality of an apparatus for collecting a biological sample, for example, adipose tissue, for subsequent reuse.
[0022] Figure 2 shows a non-restrictive modality of a filter device, with a piston arrangement manually operated.
[0023] Figure 3 shows a non-restrictive modality of a plunger arrangement with a frictional interface.
[0024] Figure 4 shows a non-restrictive modality of an apparatus for contacting biological material (for example, cells or adipose tissue), with a membrane stabilizing agent.
[0025] Figure 5 shows a non-restrictive modality of an apparatus for contacting biological material, with a retaining matrix comprising an absorbent material.
[0026] Figure 6 shows a non-restrictive modality of an apparatus to transplant biological material in an individual.
[0027] Figure 7 shows a non-restrictive modality of an apparatus for processing biological material, which comprises an arrangement with two plungers, to vary the effective volume in the apparatus chamber, to facilitate filtration and / or to allow the mixing of different agents with biological material.
[0028] Figure 8A shows a non-restrictive modality of a liposuction product processing system.
[0029] Figure 8B shows a non-restrictive modality of a liposuction product processing system.
[0030] Figure 8C shows a non-restrictive modality of a liposuction product processing system.
[0031] Figure 8D shows a non-restrictive modality of an inlet cap of a liposuction product processing chamber.
[0032] Figure 8E shows a non-restrictive modality of an outlet cover of a liposuction product processing chamber.
[0033] Figure 8F shows a non-restrictive modality of a cross-sectional view of a liposuction product processing chamber.
[0034] Figure 9A shows a non-restrictive modality of a multi-stage liposuction product processing system.
[0035] Figure 9B shows a non-restrictive modality of a multi-stage liposuction product processing system.
[0036] Figure 9C shows a non-restrictive modality of a liposuction product processing chamber.
[0037] Figure 9D shows a non-restrictive modality of a liposuction product processing chamber, with a retention matrix, with a lipophilic region and a hydrophilic region.
[0038] Figure 10A shows a non-restrictive modality of an apparatus for injecting biological material into an individual.
[0039] Figure 10B shows a non-restrictive modality of an apparatus for injecting biological material into an individual.
[0040] Figure 10C shows a non-restrictive modality of an apparatus to inject biological material into an individual.
[0041] Figure 11 shows a non-restrictive embodiment of a pressure gauge arrangement for the suction syringe.
[0042] Figure 12 shows a non-restrictive modality of an injection syringe with a 16 gauge angiocatheter manometer arrangement.
[0043] Figure 13 shows a pressure graph of the suction syringe.
[0044] Figure 14 shows pressure curves of the suction syringe.
[0045] Figure 15 demonstrates the effects of aspiration pressure on lobe weights.
[0046] Figure 16 demonstrates the effect of aspiration pressure on the histology of adipose tissue.
[0047] Figure 17 demonstrates the effect of aspiration pressure on histology scores of adipose tissue.
[0048] Figure 18 shows the effect of injection pressure readings of adipose tissue injected through a catheter or syringe.
[0049] Figure 19 demonstrates the effects of injection pressure on lobe weights.
[0050] Figure 20 demonstrates the effect of injection pressure on the histology of adipose tissue.
[0051] Figure 21 demonstrates the effect of injection pressure on adipose tissue histology scores. DETAILED DESCRIPTION OF CERTAIN MODALITIES OF THE INVENTION
[0052] The present invention relates to methods, systems and devices to improve the quality and / or viability of biological material, such as collected adipose tissue, liposuction product, adipose cells, basal cells or other cells, tissues or biological components .
[0053] Typically, the processed biological material is used for fat transplantation, both for cosmetic, reconstructive and therapeutic purposes. Biological material, for example, can be used in an autologous fat transfer procedure, that is, where material is taken from the same individual to whom it is later transferred back. In some embodiments, the quality and / or viability of the biological material is improved by treating the biological material with membrane repair / stabilizing agents or the like and / or by removing undesirable components of the biological material, such as blood cells, lipids free, excess fluid and / or excess MSA. The biological material can be processed using a system or apparatus according to the invention, which comprises one or more stages to remove one or more undesirable components (for example, free lipids, tumescent fluid, cell fragments) from the biological material. It has been found that the removal of these components improves the quality and / or viability of biological material, particularly for use as a tissue graft. The present invention also relates to methods, systems and apparatus for transplanting processed biological material, such as adipose tissue. Methods, systems and devices are designed to control one or more conditions (for example, pressure, speed or shear load), associated with the processing or injection of biological material (for example, adipose tissue) in a tissue grafting procedure and result in in improved graft quality.
[0054] The biological material processed by an apparatus or system of the invention is often the product of liposuction or adipose tissue obtained from an individual. Typically, the processed biological material comprises adipose tissue or a component thereof, including adipocytes, adipogenic cells, mesenchymal cells and / or basal cells. In general, processed biological material is biological material from which one or more undesirable components have been removed. The components can be removed by being retained in the retention matrix, passing through the filter, being linked by an antigen binding agent, removed by an antioxidant or a combination of them. Apparatus for Processing Biological Materials
[0055] Apparatus for processing biological materials are provided for in certain aspects of the invention. These devices are useful for, among other things, processing biological tissue (for example, adipose tissue) for subsequent reuse. Typically, the devices are sterile and aseptic, and suitable for use in surgical procedures. The chamber and other components of the device are often substantially sealed to prevent or minimize contamination of biological material. The apparatus of the invention generally includes a chamber to accommodate a biological material and one or more components to remove unwanted agents or to add agents (for example, MSAs) to the biological material. Therefore, the processing of a biological material can occur through one or more stages within the chamber.
[0056] Often, the camera has at least one entrance and at least one exit. Typically, at least one inlet offers a passage for biological material to enter the chamber and at least one outlet offers a passage for biological material to exit the chamber. A pressure generating device can also be included in the chamber or connected to it. The pressure generating device is typically configured and arranged to transfer biological material into the chamber through an inlet, and to discharge processed biological material out of the chamber through an outlet.
[0057] The apparatus may include a chamber that is cylindrical in shape (with a circular cross section), although other shapes may be used. The chamber may be provided with a threaded section or other coupling arrangement at either or both ends, to fix various components in the chamber typically, components that facilitate the processing of biological material present within the chamber. Other coupling arrangements, which can be used, include, for example, pressure adjustment connectors, clamps or the like. O-rings or other sealing arrangements may also be provided to form a leak-proof and / or pressure-resistant seal between each end of the chamber and an end piece (for example, a screw cap), which is configured to be connected removably at the end of the chamber. Optionally, these end pieces can be permanently attached to one end of the chamber and / or configured to be attached to other end pieces. In one configuration, a collecting cap may be provided, which allows a chamber to function as a collecting container for biological material (for example, liposuction product).
[0058] The chamber of an apparatus according to the invention may be provided in any size variation, depending on the volume of biological material (for example, adipose tissue) to be extracted, stored, processed and / or treated therein. For example, the volume of the chamber can vary from a few cubic centimeters (cc's) to about 1000 cc's or more, depending on the volume of biological material (for example, adipose tissue) to be collected and / or treated. For example, in certain embodiments, the volume of the chamber is about two to three times the volume of the adipose tissue to be treated. This excess volume allows the introduction and mixing of membrane repair or cell preservation agents with the biological material (for example, adipose tissue) in the chamber, as described in the present. The chamber can optionally include volumetric markings to indicate the amount of material contained therein. These markings can be used to determine the appropriate amount of a solution containing an MSA, cell protector or other substance to be added to the adipose tissue for treatment or processing, as described herein. In some configurations, the apparatus comprises multiple chambers, for example, 1, 2, 3, 4 or more chambers.
[0059] The camera can be equipped with any aspect ratio, for example, height to width ratio, length to width ratio. For example, the height or length of the chamber may be greater than, less than or approximately the same size as the width of the chamber. Different aspect ratios can offer certain advantages in different modalities. For example, in certain configurations, a chamber that is wider than tall (or long) can accommodate a larger filter element for a specific volume of biological tissue to be processed in the chamber. In certain configurations, the camera may have an aspect ratio (height to width, or length to width) of 1: 1, 2: 1, 3: 1, 4: 1, 5: 1 or 10D1. The chamber can have an internal diameter of cross section in a range of 0.5 cm to 1 cm, 0.5 cm to 2 cm, 0.5 to 5 cm, 1 cm to 2 cm, 1 cm to 5 cm, 1 cm to 10 cm or more. The chamber may have an internal cross-sectional diameter of about 0.5 cm, about 1 cm, about 2 cm, about 3 cm, about 4 cm, about 5 cm, about 6 cm, about 7 cm , about 8 cm, about 9 cm, about 10 cm or more. The chamber can have a length in a range of 1 cm to 5 cm, 5 cm to 5 cm, 2 cm to 10 cm, 5 cm to 10 cm, 5 cm to 20 cm, 5 cm to 30 cm or more. The chamber can be about 0.5 cm long, about 1 cm, about 2 cm, about 3 cm, about 4 cm, about 5 cm, about 6 cm, about 7 cm, about 8 cm, about 9 cm, about 10 cm, about 15 cm, about 20 cm, about 25 cm, about 30 cm, or more.
[0060] In certain configurations, a biological material (for example, adipose tissue or a component thereof, such as adipose cells (adipocytes) is kept within a single chamber, while it is being processed and each end of the chamber is provided with a threaded coupling or other connection arrangement to fix several end pieces in. For example, the chamber can be provided with an end piece in the form of an impermeable cover at a distal end, to form a container. Collected adipose tissue can be placed in the chamber for further processing using any appropriate technique. In certain configurations, a collecting cap, which includes a hose or tube, is attached to a proximal end of the chamber. The collecting cap is in communication, for example, with a hose - possession, so that the biological material (for example, liposuction product) is directed into the chamber while it is being collected. or eliminating the need to transfer biological material between multiple receptacles or containers reduces the amount of mechanical damage suffered, increases the efficiency of the process, reduces the loss of material and / or reduces the likelihood of contamination.
[0061] The chamber of an apparatus according to the invention can be configured to be used in a centrifuge. In certain configurations, fractions of a biological material in the chamber, which are separated by centrifugation (for example, oil, anti-floating) can be easily removed from the chamber. End pieces (for example, solid lids) can be attached to one or both ends of a chamber so that the lid can be placed in a centrifuge and rotated to separate fractions of biological material present in the chamber. After the separation and fractions of a biological material by centrifugation, a terminal piece containing an arrangement and filter, a retaining matrix or other device, can be attached to one end of the chamber with separate components, such as, for example, fluids or lipids free. A terminal piece that includes a pressure supply arrangement (for example, a plunger or a gas hose) can be attached to the other end of the chamber, where it can be used (for example, the plunger can be lowered or the gas hose can pressurized) to drive the separate fractions into the filter, retaining matrix or other device.
[0062] A terminal piece may be provided, which includes a container configured to be fixable in the chamber, and another plunger at a distal end of the container. The other plunger can be configured to vary the effective volume inside the chamber when the plunger is moved.
[0063] The other plunger may be formed as part of the chamber or may be provided as a removable end piece, which may be attached to one end of the chamber. Filters
[0064] Apparatus according to the present invention can be used to remove impurities from adipose tissue or other biological material placed in the chamber, using mechanical filtration and / or other processes. Impurities can include, for example, excess fluids, free lipids, blood cells, cell fragments, extracellular material and excessive amounts of certain agents or solutions, which can be added to tissue or biological material to obtain certain effects. To facilitate the removal of these impurities, the apparatus typically comprises at least one filter. The filter is configured and arranged to contact the mb, when present in the chamber of an apparatus, so that a fraction of waste of biological material passes through the filter. At least one chamber outlet is configured and arranged, in general, to allow discharge of the chamber of biological material that does not pass through the filter. Multiple filters may be provided to allow filtration of biological material in stages. The device may include 1 filter, 2 filters, 3 filters, 4 filters, 5 filters or more.
[0065] Often, a waste outlet line is provided, which is fluidly connected to the chamber, so that the fraction of waste coming out of the filter (the filtration product) is discharged through the waste outlet line. In some cases, the waste outlet line is configured and arranged to allow a stream into the chamber through the filter, to make cleaning the filter easier. Enabling the reverse flow, the material that was deposited on the filter, which results in the filter clogging, can be removed. This allows cleaning of the filter, without disassembling the chamber. Although, in some cases, the filter can be cleaned by removing it from the chamber and washing it. Therefore, in some configurations, the filter is removable and replaceable.
[0066] The filter can have a variety of shapes and sizes. The filter can be formed as a disc, an annular ring, a cylinder, a hollow cylinder, a leaf, etc. The filter can have an effective pore size in the range of about 1 pm to about 5 µm, about 1 pm to about 1 mm, about 1 to about 100 pm, about 1 pm to about 50 pm, about 10 um to about 50 pm, about 20 pm to about 50 pm, about 50 pm to about 500 pm, about 100 pm to about 500 pm, or about 100 pm to about 1 mm. The filter can have an effective pore size of about 1 pm, about 5 pm, about 10 pm, about 20 pm, about 50 pm, about 100 pm, about 250 pm, about 500 pm, about 1 mm, about 2.5 mm, about 5 mm, or more.
[0067] The filter can have an effective pore size that is relatively homogeneous across the filter. Alternatively, the filter can have an effective pore size, which is position dependent. For example, the filter may have an effective pore size in an upstream position in the chamber, which is relatively thick, and an effective pore size in an upstream position in the chamber, which is relatively thin. Thus, upstream, relatively large impurities in the biological material can pass through the filter and downstream, only relatively fine impurities in the biological material can pass through the filter. For example, the filter may have an effective pore size in a chamber position, which is in a range from 50 pm to about 100 pm, and an effective pore size in a downstream position in the chamber, which is in a range from about 1 pm to about 50 pm.
[0068] In certain modalities, in which adipose tissue is the biological material, the characteristics of the filter are selected to retain adipose cells in the chamber and allow minor liquids and impurities to pass through the filter when pressure is applied to the adipose tissue in the chamber. For example, the size of a typical fat cell is between about 60 and about 100 microns. Consequently, in certain embodiments, the effective size of the pores or passages in the filter is between about 20 microns and about 50 microns. These pore sizes allow liquids and small impurities, such as blood cells, to pass through the filter and be removed from the adipose tissue, while retaining the adipose cells and adipose tissue globules in the chamber. Filters with other effective proo sizes can also be used, based on the relative sizes of adipose cells and specific impurities or agents to be removed, where the proo size is preferably smaller than the average or minimum adip cell size and larger than the size of the impurities to be removed.
[0069] The filter can be composed of one or more of a variety of biocompatible materials. For example, the filter may be composed of one or more of ceramic, glass, silicon, stainless steel, a cobalt-chromium alloy, a titanium alloy, polytetrafluoroethylene, polypropylene and other polymers. The filter can also be coated with a material to prevent or minimize the adhesion of the components present in the biological material in the filter, which can lead to clogging of the filter.
[0070] Typically, at least one chamber outlet is configured and arranged to allow the chamber to discharge the fraction of biological material waste, after the filter outlet. When the biological material is a product of liposuction, the waste comprises at least one of lipids, blood components, tumescent fluid, individual cells and cellular fragments.
[0071] Filters or agents that are coated or treated with adsorbent materials can also be provided to remove impurities. For example, a lipophilic substance may be expected to adsorb free lipids, or surplus MSAs or cell protectors that are lipophilic, when used, from adipose tissue. These adsorbent agents can be predicted on a filter. The adsorbent materials or agents can be used instead of or in addition to absorbent materials, as described herein. Exemplified adsorbent and / or absorbent agents that can be used according to the embodiment of the present invention include, but are not limited to, hydrogels, such as polysaccharides (e.g., agarose or carboxymethyl cellulose), cross-linked PEGs, polyvinyl alcohols or copolymers thereof, polyacrylamides, polyacrylonitriles, polyacrylates and / or copolymers thereof. Pressure generating devices
[0072] As described herein, one or more pressure generating devices may be included in or connected to the chamber of a processing apparatus and biological material according to the invention. Pressure generating devices are typically configured and arranged to transfer biological material into the chamber through an inlet, and / or to discharge processed biological material out of the chamber through an outlet.
[0073] In some configurations, a pressure generating device is a pump. The pump can be configured and arranged to transfer biological material to the chamber and discharge biological material through the outlet. The device, in these configurations, typically also includes a pressure sensor, which is fluidly connected to the chamber and which includes an electrical outlet connected to a controller input. The pressure sensor provides an electrical signal to the controller, indicating a pressure detected in the chamber. The controller transmits control signals to the pump, based on the pressure detected.
[0074] In certain configurations, a terminal piece for a chamber includes a manually operated plunger mechanism, which functions as a pressure generating device. The plunger mechanism may include a rod attached to a piston body, in which the perimeter of the piston body substantially corresponds to the inner surface of the chamber, similar to the structure of a conventional syringe. Compression on the plunger increases the pressure in the chamber, and lifting the plunger reduces the pressure in the chamber. When one or more openings are provided at the lower end of the chamber, as described herein, compressing or pulling up the plunger, in each case, forces material out of the chamber or pulls material into it. Plunger pressure can be used to force impurities (for example, extracellular fluid, proteins, lipids, nucleic acids, red blood cells) through a filter, while retaining other components of biological material (for example, larger fat cells or fat globules in the chamber).
[0075] In some configurations, a force mimicking plunger arrangement is used. This arrangement and plunger includes a rod configured to fit into an opening that extends at least partially along a longitudinal axis of a sleeve. A piston body is attached to a distal end of the sleeve, or it may be formed as a part in a piece with the sleeve. The rod and the sleeve can have circular cross-sectional shapes, or other cross-sectional shapes can be used (for example, hexagonal, octagonal, square or triangular shapes). A clamping device, for example, a frictional interface, may be provided between the outer surface of the rod and the inner surface of the sleeve, when the rod is partially inserted into the sleeve. In operation, pushing the rod down with relatively small force allows the rod to frictionally grip the surrounding part of the sleeve and transmit the force to the piston body. This operation can be similar to that of a conventional syringe where applying force to the proximal end of the plunger causes the piston body to apply pressure to any substance present in the chamber. However, if the force applied to the rod exceeds a predetermined limit, the frictional interface can be configured to allow the rod to slide in relation to the sleeve, so that the rod further enters the sleeve and no additional force is transmitted to the piston body. In this way, the amount of force transferred to the piston head - and therefore the pressure applied to a material in the chamber to which the plunger is attached - is limited. The limiting force or pressure value can be determined based on the characteristics of the frictional interface between the rod and the sleeve and the piston body size. In this way, a simple force-limiting "clamping" mechanism can be provided in the plunger arrangement to avoid application and excessive force or pressure using the plunger. The maximum desired pressure or force is typically selected to avoid or reduce the likelihood of causing damage to fat cells or other cells in the chamber, when the plunger is compressed downward, for example, to force some impurities through a filter at the distal end of the chamber.
[0076] In certain configurations, a filter arrangement can be fixed at the distal end of the chamber, optionally with a waste container. A controlled pressure source, such as a pressurized gas (for example, air, N2), with a pressure regulator, may be provided in communication with the hose of the collection cap. In this way, the pressurized gas is used to increase the pressure inside the chamber in a controlled manner and to facilitate the expulsion of impurities through the filter, while avoiding the application of excessive pressure that can damage fat cells. In another embodiment, the gas may include components, such as oxygen, which can further improve the viability of the fat cells being treated.
[0077] In another embodiment, a terminal piece is provided for the chamber, which includes a filter or barrier that is arranged to provide an osmotic gradient over the filter / barrier, so that the fluid flows from the chamber to a waste container or vice versa, in response to an osmotic pressure gradient, instead of or in addition to mechanical pressure. Retention Matrices
[0078] Typically, the chamber accommodates one or more retention matrices, which are configured and arranged to contact biological material, when present in the chamber, so that a fraction of the biological material is retained in the retention matrix. Retention matrices are usually composed of one or more materials that retain components of the biological material. In this way, a retention matrix provides a mechanism for removing one or more undesirable components from biological material. Typically, at least one chamber outlet is configured and arranged to allow discharge of the biological material chamber, which is not retained in the retention matrix.
[0079] A retention matrix can be composed of a lipophilic matrix (or hydrophilic matrix), which retains lipids from biological material. The retention of the lipid fraction can occur through the adsorption of lipids in the lipophilic matrix, absorption of the lipid fraction in the lipophilic matrix, or a combination of these retention mechanisms, depending on the nature of the matrix and the composition of the lipids present in the biological material. For example, certain lipids can be adsorbed on the matrix; while certain other lipids can be absorbed by the matrix. Some lipids can be retained by either adsorption or absorption. Lipids can comprise free lipids, phospholipids, sterols, fat-soluble vitamins, fatty acids, monoglycerides, diglycerides, triglycerides, or any other components of lipids present or normally found in biological materials, such as adipose tissue. Additive components (for example, surplus additives) (which may or may not be the lipids themselves) can also adsorb or be absorbed by a lipophilic matrix. In some examples, the lipophilic matrix comprises a polysaccharide, a polyethylene glycol. crosslinked, a polyvinyl alcohol or copolymer thereof, a polyacrylamide, a polyacrylonitrile, a polyacrylate or a copolymer thereof.
[0080] The retention matrix can be composed of a hydrophilic matrix, which absorbs a hydrogel. The hydrophilic matrix can comprise a non-toxic osmotic material. The hydrophilic matrix can comprise a hyaluronic acid, a carbohydrate, gelatin, alginate, methyl cellulose or hydroxymethyl cellulose.
[0081] The retention characteristics of the retention matrix may be dependent on the position in the chamber. For example, the retention matrix can be lipophilic in an upstream position in the chamber and hydrophilic in a downstream position in the chamber. Alternatively, the retention matrix can be hydrophilic in an upstream position in the chamber and lipophilic (for example, hydrophobic) in a downstream position in the chamber. In this way, a retention matrix is able to retain different types of components of a biological matrix. In some cases, the retention matrix may comprise a relatively homogeneous distribution of different retention regions, for example, a homogeneous distribution of lipophilic and hydrophilic regions. Multiple retaining matrices may be present at different positions within the chamber. Retention matrices can retain different components of biological material, including, for example, H2O, lipids, metals, MSA, blood cells.
[0082] Any device or chamber may be disposable, including the retaining matrix and other components of the chamber. In some cases, the chamber and the system are arranged and configured to be easily disassembled into one or more components. In this way, the system can be easily cleaned and system components can be easily replaced or reassembled after cleaning. Consequently, the retaining matrix can be a component in a single piece of the chamber, which is not easily removable. Alternatively, the retaining matrix can be easily removed and replaced.
[0083] Through the use of retention matrices, impurities, including excess MSA or cell protector, when used, can be removed from the adipose tissue in the chamber. This can be achieved, for example, by contacting an absorbent material and / or an adsorbent material from a retention matrix with the adipose tissue. For example, a water-absorbent material (for example, hydrogel) can be provided in a retaining cap, which can be attached to one end of the chamber, as shown. The chamber can then remain at rest for a period of time or be gently shaken or shaken to provide sufficient contact between the adipose tissue and the absorbent material. After this contact has taken place for a sufficient time, for example, up to 5 minutes, up to 1 minute, the retaining cap with the absorbent material can be removed from the chamber, along with any absorbed water or other impurities that are present in the absorbed material. After this process, there are less impurities remaining with the fat cells in the chamber.
[0084] In certain embodiments, a container, which houses a retention matrix, can be fixed at one end of the chamber, to form a larger container to retain impurities in the biological material. A filter can be used in conjunction with the retention matrix. The filter and the retaining matrix can optionally be provided as separate end pieces, which can be attached to each other and to the chamber, or they can be provided as a single end piece, which is fixed to the chamber. The biological material with impurities (including any MSA or cell protector, when used), can then be contacted with the retention matrix, for example, by inverting the chamber / absorbent container assembly. A relatively large retention surface area provides improved contact between the biological material and the absorbent material, to facilitate better absorption of impurities.
[0085] The chamber can also include one or more binding agents and antigen, to bind and separate target antigens present in biological material. Antigen binding agents are typically immobilized on a solid support. In some configurations, the antigen binding agent is immobilized on a retention matrix (for example, a lipophilic matrix or hydrophilic matrix). In other configurations, the antigen binding agent is immobilized on a separate structure (for example, a chamber surface) or matrix. The antigen binding agent can specifically bind to a cell surface molecule, extracellular matrix molecule or other target. The cell surface molecule may be present in a red blood cell, white blood cell, platelet, stromal cell, bacteria, virus or other target. In general, the target of the antigen binding agent is an undesirable component of biological material. The antigen binding agent is, in general, an antibody or an antigen binding fragment thereof.
[0086] The chamber can also include one or more antioxidants to remove free radicals from biological material. Antioxidants can be immobilized on a solid support in the chamber. In some configurations, the antioxidant is immobilized on a retention matrix (for example, a lipophilic matrix or hydrophilic matrix). In other configurations, the antigen binding agent is immobilized on a separate structure (for example, a chamber surface) or matrix. Examples of antioxidants useful in the present invention include, but are not limited to, glutathione, vitamin C, vitamin E or an enzyme, such as a catalase, superoxide dismutase or peroxidase. Additive Reservoir
[0087] The system may also comprise a reservoir configured and arranged to accommodate an additive solution. The additive solution can comprise a membrane stabilizing agent (MSA), a growth factor, an antioxidant or other suitable additive described herein or otherwise known in the art. The MSA can be a triblock copolymer of the form: polyethylene glycol-polypropylene glycol-polyethylene glycol, such as Poloxamer-P188. The reservoir is typically configured and arranged to allow transfer of the additive solution from the reservoir to the biological material, before the biological material contacts the retention matrix. However, the reservoir can be configured and arranged to allow transfer of the additive solution to the biological material at any stage of processing in the chamber. Typically, the additive solution is transferred from the reservoir to the chamber using a pump or similar device. In some cases, the retention matrix is configured and arranged to retain an excess of one or more components of the additive solution by adsorption or absorption.
[0088] Damage to the collected cells (for example, adipocytes or similar), when present, can be repaired or avoided by mixing the adipose tissue with a membrane stabilizing agent (MSA) or cell protector, such as, for example, Poloxamer P188 or lipoic acid. MSAs or cell protectors that can be used to improve cell viability are described, for example, in International Application No. PCT / US2009 / 005727, filed October 21, 2009 and US Patent Application Publication No. 2010/0104542 . The MSA or cell protector can be inserted into the chamber in several ways. For example, the MSA or cell protector can be supplied in a solution that is previously charged in the chamber or is added to a solution container in communication with the chamber. Certain MSAs or cell protectors can also be provided, for example, in the form of a solid, such as a powder, or in liquid form (i.e., not in solution). The MSA or cell protector is preferably sterile to prevent contamination of the cells or tissue being treated.
[0089] In certain modalities, a cover is provided at the proximal end of the chamber, to facilitate mixing of the MSA or cell protector with the biological material. The MSA solution or cell protector can be introduced into the chamber and mixed with the biological material in it by inverting the solution container / chamber set, so that the MSA solution or cell protector passes through the filter and into the chamber. Alternatively, a plunger mechanism can be attached to the proximal end of the chamber, so that the plunger can be partially removed from the chamber to pull the MSA and cell protector solution through the filter and into the chamber. Other techniques can also be used to introduce the MSA solution or cell protector into the chamber. For example, a terminal piece provided at one end of the chamber can be removed and a specific amount of MSA solution or cell protector can be poured into the chamber.
[0090] The MSA solution or cell protector can be supplied in a previously measured amount, for example, if P188 is used as the MSA, it can be supplied in a 10 mg / ml aqueous solution (for example, a solution of PBS or other buffered solution, comprising MSA), in which the volume of solution used is approximately the same as the volume of adipose tissue to be treated The mixture of adipose tissue and MSAS or cell protector in the chamber can be gently shaken, subsequently mixed by inversion, spin etc. and / or left to rest for a sufficient duration to allow the MSA or cell protector to interact with the cells. For example, the mixture can be left to stand for about 1 minute, about 5 minutes, about 10 minutes, about 20 minutes, or for longer. Shorter lengths of time can be used. In some cases, the mixture is agitated to perfect the mixture of the cells in the biological material and the MSA or cell protector.
[0091] In certain modalities, MAS, cell protector and / or other treatment / processing substance is present at a temperature below normal body temperature and / or below the temperature of the neighborhood ("room temperature"). Alternatively or additionally, the cell-containing chamber is cooled before and / or after the introduction of adipose tissue or other cells. These refrigeration temperatures can also facilitate the conservation and viability of the cells or tissue being processed.
[0092] After the adipose tissue has been sufficiently mixed or exposed to the MSA or cell protector, the MSA or cell protector is optionally removed from the biological material. Materials other than or instead of MSAs or cell protectors, for example, any substance that can have a beneficial effect on cells or other biological material in the chamber, can also be used as described herein. These materials and / or other cell protectors or MSAs can be used in combination or can be used to sequentially treat the collected cells or tissue, according to the apparatus and techniques according to the invention described herein. Agitation
[0093] The system may also be equipped with a stirring device, configured and arranged to stir biological material, while present in the chamber. In some configurations, the agitation of the biological material, both in the chamber and before entering the chamber, facilitates the movement of the biological material into and out of the chamber, as well as to contact the biological material with retaining matrices, filters and other components chamber, to facilitate the removal of one or more undesirable components of biological material. The stirring device can be an internal stirring or mixing mechanism, which is in direct or indirect contact with the biological material. The stirring device can also be an external device, such as a vibrating device, oscillating table or other similar device, suitable for stirring the biological material externally. Methods for Processing Biological Materials
[0094] Methods for processing biological materials are provided for in the present. The methods are particularly useful for fat transplant procedures. For example, the liposuction product obtained from an individual can be processed using the methods and to produce adipose tissue suitable for transplantation in the individual for cosmetic or reconstructive purposes. The methods, which are often used to prepare tissue grafting, typically involve obtaining an appropriate apparatus as described herein; obtaining biological material (typically from an individual); and causing the pressure generating device to transfer biological material into the apparatus chamber through the chamber inlet and discharge biological material out of the chamber through the chamber outlet. In some configurations, the pressure generating device is not required to load biological material into the device. In these configurations, biological material can be added manually (for example, dumped) in the chamber. The pressure generating device in this configuration would facilitate the movement of biological material through one or more stages of processing the device and, optionally, the discharge of the processed biological material through the outlet.
[0095] In certain modalities, the liposuction product is subjected to centrifugation, before common processing using the device. In some cases, after centrifugation, an oil layer and / or an infraflutuating layer is removed from the liposuction product.
[0096] In some device configurations, any combination of terminal parts and procedures described herein can be used to collect, treat and / or process biological materials (for example, adipose tissue). Individual procedures can also be performed in different orders and / or more than one, such as using a plurality of absorbent covers with fresh absorbent materials. The type and number of processes to be used can be based on several factors, such as the amount of adipose tissue being treated, the desired level of purification of the adipose cells, the size of the membrane repair or other treatment of the fat cells, which is desired or selected, the location from which the cells or tissue were extracted, the location of the transplant etc.
[0097] After the adipose tissue has been treated using any of the various procedures and devices described herein, a terminal piece, which includes a needle, catheter, cannula or other appropriate opening, can be attached to a distal end of the chamber to supply the processed biological material to an individual. The needle, catheter, cannula or other appropriate opening can be used to facilitate the transplantation of adipose tissue at one or more specific locations on a patient's body. For example, a plunger mechanism can be attached to the proximal end of the chamber and used to force treated and / or processed fat cells through the needle, catheter, cannula or other appropriate opening. In this way, the collected adipose tissue can be treated and transplanted using a single chamber, which can reduce the amount of damage to adipose cells and improve their total viability. Apparatus for Injecting, Transplanting or Transferring Biological Materials
[0098] Devices are provided for injecting, transplanting or transferring biological materials under conditions that result in improved quality, consistency and viability of the biological material for grafting purposes, compared to existing devices. The apparatus typically comprises a chamber with at least one outlet; and a pressure generating device, configured and arranged to generate positive pressure within the chamber, which is at or below a predetermined threshold, where the positive pressure is sufficient to cause a biological material, when present in the chamber, to be discharged through the exit. The device can include a pressure sensor, configured and arranged to measure the pressure inside the chamber.
[0099] In general, the predetermined threshold is a pressure above which the biological material has a relatively low viability as a tissue graft, after discharge from the outlet to a graft site in an individual. The size of the viability can be compared with a control graft sample with a set of desired viability conditions, or a historical value or values, which represent a desired set of viability conditions. Viability can be assessed using methods well known in the art. For example, viability can be determined by assessing the extent of graft retention (for example, by thickness, volume, adipocyte content, cell viability or other parameter of no inflammation) and / or growth after transplantation, in a or more times after the transplant. The viability of the graft can be determined qualitatively by evaluating the aesthetic appearance of an individual who received a fat graft. Clinically, graft viability can also be determined using volume measurements by MRI or 3D images. Before fat grafting, viability can be measured using an automatic cell counter. Again, the determination can be performed one or more times after the graft. For example, when the graft was made to reduce or eliminate one or more facial wrinkles, preservation and an apparent reduction or absence of wrinkles on one or more occasions after the graft may be indicative of sufficient viability. The thickness of a graft and quality can also be determined by other quantitative methods, such as using ultrasound or other means to determine the quality of a graft over time in an individual. The experienced technician is familiar with other methods that can be used to assess the viability of the graft.
[00100] The predetermined threshold can be about 2 atm, about 3 atm, about 4 atm, about 5 atm, about 6 atm, or more. The predetermined threshold can be in the range of about 2 atm to about 3 atm, about 3 atm to about 4 atm, about 2 tom to about 5 atm, about 2 tom to about 6 atm, or about from 4 atm to about 6 atm. Often, the predetermined threshold is a pressure, above which the speed of the biological material being discharged from the outlet exceeds a predetermined maximum. For example, the predetermined maximum can be about 5 cm / s, about 10 cm / s, about 20 cm / s, about 30 cm / s, about 40 cm / s, about 50 cm / s , about 60 cm / s, about 70 cm / s, about 80 cm / s, about 90 cm / s, about 100 cm / s, about 150 cm / s, about 200 cm / s, about 250 cm / s or more. The predetermined maximum can be in the range of about 5 cm / s to about 20 cm / s, about 10 cm / s to about 50 cm / s, about 20 cm / s to about 100 cm / s, about 50 cm / s about 200 cm / s, about 50 cm / s about 250 cm / s, or more. The predetermined maximum can be about 265 cm / s. The positive pressure is often maintained in such a way that the speed of the biological material being discharged from the outlet is in the range of about 5 cm / s to about 265 cm / s.
[00101] In some configurations, the outlet is positioned at a distal end of the chamber, the chamber comprises an opening at a proximal end, and the pressure generating device is a plunger arrangement, configured and arranged to pass through the opening at the end proximal and to be movably arranged within the chamber, so that a displacement of the plunger arrangement within the chamber towards the distal end generates the positive pressure necessary to discharge the material into the chamber. In some configurations, the plunger arrangement can be advanced into the chamber by compressing the plunger arrangement (for example, manually). The plunger arrangement can also be operated automatically using an electric motor, for example, which is coupled to the plunger arrangement, so that rotation of the motor causes the plunger arrangement to advance into or out of the chamber. The operation of the motor can be controlled automatically based on a pressure detected in the chamber, to ensure that the pressure in the chamber does not exceed the predetermined threshold.
[00102] In some configurations, the plunger arrangement comprises a force limiting clamping arrangement, which couples a rod to a piston. The force limiting clamping arrangement may comprise a frictional interface, which mechanically attaches rod to the piston. The force limiting clamping arrangement may be configured and arranged to limit the maximum developed pressure in the chamber to the predetermined threshold, compressing the rod of the piston arrangement towards the distal end.
[00103] The arrangement may comprise a piston displacement device (for example, an electric motor, compressed air drive, vacuum drive) configured and arranged to move the plunger arrangement towards the distal end. The plunger arrangement may be configured and arranged to move the plunger arrangement towards the distal end, at a predetermined speed. The predetermined speed can be a displacement speed of the plunger arrangement, which results in a positive pressure in the chamber at or below the predetermined threshold, while discharging the biological material from the outlet. The apparatus may also include a controller configured and arranged to generate control signals, which activate the plunger displacement device to move the plunger arrangement towards the distal end. The device may also include a pressure sensor, which is in fluid communication with the chamber and which includes an electrical outlet connected to an input of the controller, so that the pressure sensor provides an electrical signal to the controller, which indicates a pressure detected in the chamber, in which the controller transmits the control signals to the plunger based on the pressure detected.
[00104] In some configurations, the pressure generating device for injecting the processed biological material is a pump. The pump can be configured and arranged to transfer biological material to the chamber and discharge biological material through the outlet. The device, in these configurations, also typically includes a controller configured and arranged to generate control signals, which activate the pump to generate positive pressure. The device can also include a pressure sensor, which is in fluid communication with the chamber and which includes an electrical outlet connected to an input of the controller, so that the pressure sensor provides an electrical signal to the controller, which indicates a pressure detected in the chamber, where the controller transmits the control signals to the plunger based on the pressure detected.
[00105] In some configurations, the chamber outlet is in fluid communication with a cannula (for example, blunt cannula) or catheter. Although any appropriate size can be used, the cannula or catheter is typically 12, 14, 15, 16, 17 or 18 caliber.
[00106] The chamber is configured and arranged, in general, to contain 1 ml to 1 liter of biological material. However, a variety of volumetric sizes can be outlined. In some configurations, the chamber is configured and arranged to contain a volume of biological material ranging from 1 ml to 1 liter, 1 ml to 500 ml, 1 ml to 100 ml, 1 ml to 50 ml, 50 ml to 100 ml, 20 ml to 100 ml, or 0.5 ml to 1 ml.
[00107] Methods for transplanting processed biological material, for example, adipose tissue, for autologous fat grafting, are also provided for. Typically, the methods involve obtaining biological material processed according to any of the methods described herein; and transplant the biological material into an individual. Method for injecting, transplanting or transferring biological materials may include obtaining any of the devices described herein; load the biological material in a device chamber; and causing the pressure generating device of the apparatus to generate a positive pressure within the chamber, which is at or below the predetermined threshold to cause the biological material to be discharged through the outlet. Often, pressure in the chamber is monitored during transplantation to ensure that the pressure does not exceed the predetermined threshold.
[00108] Typically, the methods involve obtaining biological material processed according to any of the methods described herein; and transplant the processed biological material into an individual. For fat transfer (grafting) procedures, the processed biological material generally comprises adipose tissue or one or more components thereof. In autologous fat transfer procedures, adipose tissue is typically obtained as a liposuction product from the individual; then it is processed and injected back into the patient in a new location. The biological material can be obtained by extracting adipose tissue from the abdomen, sew, flank region or gluteal region of the individual or by extraction of liposuction product, which comprises this individual's adipose tissue. For cosmetic purposes, processed biological material is often transplanted under the skin of a nasolabial fold, nasojalal region, malar, chin, forehead, lower eyelid or upper eyelid of the individual.
[00109] Although the exemplified methods and apparatus described herein are described, in most cases, with respect to the treatment or processing of adipose tissue (for example, collected fat), they can be used with any biological material that benefits from any one of the exemplified processing procedures described herein. These procedures include, for example, removal of impurities or certain components by filtration and / or absorption / adsorption. The biological material (for example, adipose tissue or other tissue) can be combined with any substance that may produce a beneficial or desirable effect on the biological material, in addition to the MSAs or cell protectors described herein. Excessive substance can also be removed after combining it with biological material using the apparatus and exemplified methods described herein. Kits
[00110] The devices and systems described in the present can be made available as a kit to treat collected adipose tissue, liposuction product or other biological material. The kit can include one or more chambers, which can be discarded and / or sterilized. These chambers can be provided in different sizes / volumes, as described herein. A variety of end pieces can be provided, which can be attached to one end of the chambers. Such end pieces may include collection lids, filters, waste containers, lids or containers provided with absorbent or adsorbent materials or agents, plunger mechanisms, needles or cannulas, etc. These terminal pieces can be provided in different sizes, which are configured to be fixed in different sized chambers. An amount of one or more MSAs or other agents to treat or process biological material can also be anticipated. These agents can be made available in a liquid form, for example, as a pure liquid, suspension, solution or emulsion, or as a crystalline solid or powder, or even in a gaseous form. These substance (s) can be supplied in a bottle or distribution container, or in one or more containers, which can be configured to be fixed in the chamber containing the adipose tissue or other biological material . The substance (for example, an MSA or cell protector) can be supplied in these containers in volumes previously measured for use with appropriate volumes of collected adipose tissue, etc.
[00111] The above merely illustrates the principles of the invention. Several modifications and combinations of the described modalities are evident for those who are versed in the technique, in view of the teachings of the present. Therefore, it should be understood that those skilled in the art are able to derive numerous techniques that, although not explicitly described in the present, incorporate the principles of the invention and, therefore, are within the spirit and scope of the invention. Exemplified embodiments of the invention are described in more detail by the following examples. These modalities are examples of the invention, which, as someone skilled in the art recognizes, is not limited to the exemplified modalities.
[00112] All references referenced herein are incorporated by reference for the purposes described herein. EXAMPLES Example: Device for collecting a biological sample, for example, adipose tissue, for subsequent reuse.
[00113] Figure 1 shows a device to collect a biological sample, for example, adipose tissue, for subsequent reuse. The apparatus includes a chamber, which is cylindrical in shape. The chamber is provided with a threaded section, but other coupling arrangements can be used at either or both ends. Other coupling arrangements, which can be used, include, for example, pressure adjustment connectors, clamps or the like. O-rings or other sealing arrangements may also be provided to form a leak-proof and / or pressure-resistant seal between each end of the chamber and an end piece (for example, a screw cap), which is configured to be connected removably to the end of it. Optionally, these end pieces can be permanently attached to one end of the chamber and / or configured to be attached to other end pieces. The chamber can optionally include volumetric markings, to indicate the amount of material contained therein, as shown in Figure 1. In this example, fat cells (adipocytes) are kept within a single chamber, while being processed. Each end of the chambers is equipped with a threaded coupling or other connection arrangement to fix several end pieces in it. For example, as shown in Figure 1, the chamber may be provided with an end piece in the form of an impermeable cover at the distal end, to form a container. Collected adipose tissue can be placed in the chamber for further processing using any appropriate technique. In certain embodiments, a collection cap, which includes a hose or tube, is attached to the proximal end of the chamber, as shown in Figure 1. Example 2: Filter device with a manually operated plunger arrangement.
[00114] In another example, a filter, as shown in Figure 2, can already be attached to the distal end of the chamber shown in Figure I. For example, the collection cap, as shown in Figure 1, when used, is removed , and a plunger mechanism is attached to the proximal end of the chamber. The chamber is then inverted, so that the distal end of the chamber is at the top, and the filter arrangement is fixed at the distal end of the chamber. In certain embodiments, a waste container is attached to the chamber, as shown in Figure 2, or the filter and waste container arrangement are provided as a single component. A terminal piece that includes a manually operated plunger mechanism is attached to the proximal end of the chamber. Example 3: Plunger arrangement with a frictional interface.
[00115] A limited force plunger arrangement, which can be used with certain embodiments of the present invention is shown in Figure 3. This plunger arrangement includes a rod configured to fit into an opening that extends, at least partially, along a longitudinal axis of a glove. A piston body is attached to a distal end of the sleeve, or it may be formed as a single piece with the sleeve. The rod and glove can have circular cross-sectional shapes, or other cross-sectional shapes can be used (for example, hexagonal, octagonal, square or triangular shapes).
[00116] A clamping arrangement, for example, a frictional or similar interface, may be provided between the outer surface of the rod and the inner surface of the sleeve, when the rod is partially inserted into the sleeve, as shown in Figure 3. operation, pushing the rod down with a relatively small force, allows the rod to fictionally cling to the surrounding part of the sleeve and transmit the force to the piston body. This operation can be similar to that of a conventional syringe, where applying force to the proximal end of the plunger causes the piston body to compress on any substance provided in the chamber, for example, as shown in Figure 2.
[00117] If the force applied to the rod exceeds a predetermined limit, the frictional interface may be configured to allow the rod to slide in relation to the sleeve, so that the rod additionally enters the sleeve and no additional force is transmitted to the piston body. In this way, the amount of force transferred to the piston head - and therefore the pressure applied to a material in the chamber to which the plunger is attached - is limited. This limiting force or pressure value can be determined based on characteristics of the frictional interface between the rod and the sleeve and the piston body size. Thus, a simple force-limiting "clamping" mechanism may be provided in the plunger arrangement, to prevent the application of excessive force or pressure using the plunger. The desired maximum force or pressure is typically selected to avoid or reduce the likelihood of causing damage to fat cells or other cells in the chamber, when the plunger is compressed downward, for example, to force some impurities through a filter at the distal end of the chamber. Example 4: Apparatus for contacting biological material (for example, cells or adipose tissue) with a membrane stabilizing agent.
[00118] Damage to the collected cells can be repaired or prevented by mixing the adipose tissue with a membrane stabilizing agent (MSA) or cell protector, such as, for example, Poloxamer P188 or lipoic acid. The MSA or cell protector can be provided in a solution that is pre-loaded in the chamber or added to a solution container in communication with the chamber. Certain MSAs or cell protectors may also be provided, for example, in the form of a solid, such as a powder, or in liquid form. In the apparatus shown in Figure 4, the upper part of the container is configured to be fixed at the distal end of the chamber, and a filter is optionally provided between the chamber and the solution container. The MSA or cell protector is sterile to prevent contamination of the cells or tissue being treated.
[00119] A cap is provided on the proximal end of the chamber, as shown in Figure 4. The MSA or cell protection solution is introduced into the chamber and mixed with the adipose tissue in it, inverting the chamber / solution container set shown in Figure 4, so that the MSA or cell protection solution passes through the filter and into the chamber. Alternatively, a plunger mechanism can be attached to the proximal end of the chamber, as shown in Figure 2. The chamber / solution container assembly is inverted and the plunger is partially removed from the chamber to propel the MSA or cell protection solution through filter and into the chamber. Example 5: Apparatus for contacting biological material with an absorbent material.
[00120] Impurities, including excess MSA or cell protector, when used, can be removed from the adipose tissue in a chamber providing an absorbent / or adsorbent material in contact with the adipose tissue. For example, a water-absorbing material may be provided in a retaining cap, which can be attached to one end of the chamber, as shown in Figure 5. The chamber can then be left to rest for a period of time or be gently shaken or agitated to provide sufficient contact between the adipose tissue and the absorbent material. After this contact has taken place for a sufficient time (for example, 3 seconds at 5 minutes), the retaining cap with the absorbent material can be removed from the chamber, along with any absorbed water or other impurities, which are present in the material absorbed. After this process, there are less impurities remaining with the fat cells in the chamber.
[00121] The lid shown in Figure 5 can be elongated to form an absorbent container, similar to the waste container shown in Figure 2. At least part of the inner surface of the absorbent container is provided with a material pad. The absorbent container can be attached to the 'distal end of the chamber to form a larger container (for example, similar to that shown in Figure 2, with or without the central filter). Alternatively, a filter is used in conjunction with the absorbent container. The filter element and the absorption container are optionally provided as separate end pieces, which can be attached to each other and to the chamber, or are provided as a single end piece, which is fixed to the chamber. The adipose material with impurities (including any MSA or cell protector, when used) must then be contacted with the absorbent material, for example, by inverting the absorbent chamber / container assembly. The relatively large area of the internal surface of the absorbent container provides improved contact between the adipose tissue and the absorbent material to facilitate better absorption and impurities of the adipose tissue. This configuration also facilitates the absorption of larger amounts of impurities (for example, fluid, large volumes of an MSA solution or cell protector) by the greater amount of absorbent material that can be made available in the absorbent container. Processed adipose tissue can be injected into a graft site in an individual using an apparatus as shown in Figure 6 or 10. Example 6: Double piston apparatus for processing biological material
[00122] Figure 7 shows an apparatus comprising a double piston arrangement, to vary the effective volume in the apparatus chamber, to facilitate filtration and / or to allow the mixing of different agents with the biological material. An end piece is provided that includes a container configured to be connected to the chamber and another plunger or other similar arrangement at a distal end of the containers (that is, at the end of the container opposite the proximal end, which is configured to be connected to the camera or another end piece). The other plunger can be configured to vary the effective volume of the container when the plunger is moved. The other plunger may be formed as part of the container or it may be provided as a removable end piece, which may be attached to one end of the chamber, similar to the upper end piece / plunger arrangement shown in Figure 7. The container may also include a filter or other restrictive but permeable barrier that is located between the container and the chamber, as shown in Figure 7.
[00123] The apparatus shown in Figure 7 can be used to facilitate the introduction and / or removal of a liquid or fluid from the container to the chamber or vice versa. For example, a first plunger located partially inside the chamber is compressed, while a second plunger partially inside the container is simultaneously removed. This "push-pull" operation can increase the pressure in the chamber and reduce the pressure inside the container. This pressure differential is used to force a quantity of fluid to pass through the filter or other barrier and into the container. Reversing the direction of the piston movements, fluid or liquid is forced into the container chamber. In certain embodiments, the movement of the two pistons is coupled, for example, connecting their respective rods with a mechanical coupling. In other embodiments, a plunger is compressed or withdrawn, while another plunger is allowed to move freely. This free movement accommodates the volume of material that passes from the container into the chamber or vice versa, when a force is applied to a single plunger.
[00124] The container can also contain an amount of an MSA, cell protector or other fluid or substance that must be added to the biological material in the chamber. The container can be provided with a filter or other permeable barrier covering at least a part of the end of the container, which is configured to be fixed in the chamber. The properties of this filter can be selected to prevent leakage or overflow of the fluid in the container under atmospheric pressure, but let the fluid be introduced into the chamber when the container is attached to one end of the chamber and a force is applied to one or both pistons.
[00125] A removable film or sheet may be provided on the filter end of the container for storage. This film can also prevent the leakage of fluid from the container / or help maintain the sterility of the filter and container contents. This film can be detached or otherwise removed just before fixing the container to the chamber. A film that can be perforated to enable the MSA to run into the biological material can also be envisaged.
[00126] The container shown in Figure 7 can be initially empty and used as a waste container for any fluids that are removed from the chamber through the filter, for example, applying a force to one or both pistons, as described herein. Example 7: Liposuction product processing system
[00127] Figure 8A shows a liposuction product processing system. This system includes a liposuction product container to collect liposuction product, which was obtained from a patient. The liposuction product enters the liposuction product container through the liposuction product inlet. The liposuction product inlet is provided with an inlet valve to fluidly separate the inlet from the liposuction product container. The liposuction product container is also in fluid communication with an additive opening that is provided with a valve to fluidly isolate the additive opening from the liposuction product container. The additive opening can be used to deliver a variety of different additives to the liposuction product, which aid in the processing of the liposuction product or which improve the viability of the processed adipose tissue. Any of the components of this system can be coupled via separable or non-separable connections.
[00128] The liposuction product container is also in fluid communication with a liposuction product processing chamber. The processing chamber comprises a container into which undesirable parts of the liposuction product are removed. Between the liposuction product container and the liposuction product processing chamber is a pressure generating device. The pressure generating device aspirates liposuction product from the liposuction product container and pushes the liposuction product into the liposuction product processing chamber through a check valve. The check valve in this configuration prevents the return of the processed liposuction product to the liposuction product container.
[00129] The production and liposuction container can comprise several of different components useful to isolate undesirable parts of the liposuction product. In some configurations, the liposuction product processing chamber includes a filter to remove blood cells, debris and cells, free lipids, extracellular material and other agents from the liposuction product. In some configurations, in order to propel the filtration product through the filter, it is convenient to close the outlet valve and operate the pressure generating device to generate a positive pressure inside the liposuction product processing chamber, which is sufficient to move the filtration product through the filter. The filtration product passes into a waste cavity and exits the processing chamber through a waste outlet valve and into a waste container. The filtration product therefore includes several undesirable parts of the liposuction product, including cell fragments, extracellular material and other agents.
[00130] The liposuction product processing chamber may also include other components, including, for example, a retention matrix, immobilized antigen binding factors or antioxidants to remove free radicals from the liposuction product.
[00131] The liposuction product processing system also includes a container of processed adipose tissue, which is a container that is used to collect adipose tissue, which remains after processing the liposuction product. Processed adipose tissue is often suitable for injection directly into the individual for cosmetic or reconstructive purposes.
[00132] Figure 8B shows another configuration of a liposuction product processing system. This configuration is equipped with a compressed air inlet, which is in fluid communication with the liposuction product processing chamber. The liposuction product inlet and the additive inlet are in fluid communication with a bladder. Therefore, the liposuction product and any additives are collected inside the bladder. The system is configured in such a way that compressed air surrounds the bladder to prevent fluid out of the liposuction product container towards the liposuction product processing container. In some configurations, compressed air enters the liposuction product processing chamber with the liposuction product inlet valve and the additive opening inlet valve closed, so that the fluid does not return through the inlet lines. . Alternatively, the inlet lines may be provided with check valves in the line, which prevent the return of the liposuction product into the inlet lines.
[00133] Figure 8C shows a cross section of an exemplary liposuction product processing chamber. The liposuction product processing chamber comprises an inlet line, which is fluidly connected with a retaining matrix. The retention matrix can include, for example, a hydrophilic material and / or lipophilic material. The retaining matrix is in contact with a surrounding filter. The filter is configured to let impurities pass through the filter and into a waste cavity. As the liposuction product moves through the chamber, from the entrance towards the exit, undesirable parts of the liposuction product are retained in the matrix. In some cases, in order to facilitate the filtration of the liposuction product, an outlet valve is closed, so that sufficient positive pressure develops within the liposuction product processing chamber, to allow passage filtration product through the filter and into the waste cavity. The filtration product in the waste cavity is removed from the liposuction product processing chamber through the waste outlet, which is present in the outlet lid.
[00134] Figures 8D and 8E show alternative views of the inlet and outlet cover and show the inlet and outlet openings of the liposuction product processing chamber.
[00135] Figure 8F shows a cross section of the liposuction product processing chamber and illustrates the presence of the retention matrix inside and surrounded by the filter component, which, in turn, is surrounded by a waste cavity. Example 8: Multi-stage liposuction product processing system.
[00136] Figure 9A shows a non-restrictive modality of a multi-stage liposuction product processing system. This system includes an additive chamber, a retention matrix, an antioxidant / antigen binding chamber and a filter chamber. The liposuction product that enters the system passes first into an additive chamber, which is in fluid communication with an additive opening to add components, such as membrane stabilizing agent, anti-oxidants and other components, which can improve the viability of the processed adipose tissue obtained from the liposuction product. The additives contact the liposuction product inside the additive chamber and surplus additive leaves the additive chamber into the waste cavity. The second stage of the system comprises a retention matrix. The retention matrix can include, for example, one or more hydrophilic or lipophilic materials to retain water and lipids (or other lipophilic molecules) from the liposuction product. The third stage of the system includes an antioxidant or antigen binding chamber. In this chamber, antioxidant and / or antigen binding factors are immobilized on a solid support and, in the case of antioxidants, separate free radicals from the liposuction product, and in the case of antigen binding agents, separate one or more antigens target of the liposuction product. The fourth stage of the liposuction product processing system includes a filter configured to remove one or more unwanted parts of the liposuction product. Undesirable parts pass through the filter and exit as a filtration product through filter waste openings into a waste cavity.
[00137] Additive and excess filtration product, present in the waste cavity, leave the system through a waste outlet opening. Processed liposuction product, which mainly comprises adipose tissue suitable for injection into an individual, leaves the liposuction product processing system through an outlet opening, which is configured with an outlet valve. Processed liposuction product can be added directly to a syringe (as shown in Figure 9A), which is suitable for injection directly into an individual. Alternatively, the processed liposuction product can be deposited in a container for later use or cryopreservation.
[00138] Figure 9B shows an alternative configuration of a multi-stage liposuction product processing system. This configuration includes the coiled tube, which passes through the various stages of the liposuction product processing system. The rolled porous tube (s) can pass through the entire system or can be present only at certain stages (for example, additive stage and filter stage). The coiled tube is of porous construction and allows the additive in the additive chamber to freely contact the liposuction product. In the second stage, the liposuction product diffuses directly into the retention matrix, in which certain undesirable parts (for example, water, free lipids) of the liposuction product are retained. Similarly, in the third stage, the liposuction product contacts immobilized antioxidants and / or immobilized antigen binding factors that facilitate the additional removal of undesirable parts of the liposuction product. In the fourth stage, the liposuction product can freely contact the filter, thereby allowing the filtration product to pass through the filter into the waste cavity.
[00139] Figure 9C shows a liposuction product processing system, in which the liposuction product that enters a processing chamber travels along multiple paths. Each path comprises a retaining matrix surrounded by a filter. Liposuction product that passes through any retention matrix has undesirable parts retained within the retention matrix. A portion of the liposuction product also passes through the filter. Typically, the liposuction product processing chamber outlet valve is closed, to facilitate pressure generation within the liposuction product, in order to prevent filtration product through the filter. Filtration product, which passes through the filter enters a waste cavity and leaves the liposuction product processing chamber through a waste outlet valve. The processed liposuction product leaves the liposuction product processing chamber through the outlet valve and can be added directly to a syringe for injection into an individual or can alternatively be deposited in a container for later use or cryopreservation.
[00140] Figure 9D shows another multipath liposuction product processing system. The chamber comprises a two-stage retention matrix, which includes a lipophilic part upstream and a hydrophilic part downstream. In the lipophilic part, lipids and other lipophilic molecules are retained in the retention matrix. In the hydrophilic part, water and other hydrophilic molecules are retained. The filtration product passes through the filter, enters the waste cavity and leaves the system through the waste outlet. The processed liposuction product can be deposited in a syringe for injection directly into an individual or into a container for later use or cryopreservation. Example 9: Apparatus for injecting biological material into an individual.
[00141] Figures 10A-1-C show several devices for injecting adipose tissue. The devices in Figure 10A are configured to accommodate adipose tissue to be injected into an individual. The apparatus includes a piston arrangement configured to generate pressure within the chamber, which pushes the tissue out of the apparatus through the outlet. The piston arrangement in this configuration is coupled with a drive motor, which functions as a piston displacement device. The drive motor rotates a drive shaft, which is coupled within a shaft cavity of the piston arrangement. Rotation of the drive shaft results in displacement of the piston arrangement towards the distal end of the device thereby reducing the volume in the chamber. The drive motor can also be turned in the opposite direction, to drag the piston arrangement towards the proximal end of the device, with which the volume of the chamber increases.
[00142] The drive motor can be powered by a battery (as shown in Figure 10A). But, alternative energy sources, including AC or AC- to-DC power supplies, can be used. The operation of the drive motor is controlled by a controller, which is electrically connected to a pressure sensor. The pressure sensor detects the pressure inside the chamber, communicates an electrical signal indicating the pressure inside the chamber to the controller. The controller then operates the drive motor in response to the detected pressure. This device is configured to ensure that the drive motor operates at such a speed that the pressure inside the chamber is within a predetermined maximum. By controlling the pressure in the chamber in this way, the speed and shear load applied to the adipose tissue when it exits through the outlet can be controlled to ensure adequate adipose tissue viability. The controller can also be configured to receive one or more user inputs. For example, a user can enter a setpoint pressure, where the controller operates the drive motor to maintain the setpoint pressure specified by the user.
[00143] Figure 10B shows an injection device equipped with a mechanically controlled piston arrangement. This piston arrangement includes a frictional interface, in which a rod is connected to a sleeve via a frictional interface. The frictional interface is arranged and configured to limit the maximum force that can be generated by compressing the rod and, therefore, to limit the maximum pressure inside the chamber. By controlling the pressure in the chamber in this way, the speed and shear load applied to the adipose tissue, when it exits through the outlet, can be controlled to ensure adequate adipose tissue viability.
[00144] Figure lOCshows an injection device equipped with a pressure sensor and a display device. The device can be operated by compressing the piston arrangement manually. The device includes a pressure sensor with a display device, so that the user can monitor the pressure developed inside the chamber and ensure that the pressure developed inside the chamber does not exceed a predetermined maximum. The predetermined maximum is, as in the apparatus shown in Figure 10B, a pressure above which undesirable speeds and shear loads are applied to the adipose tissue when it exits the chamber outlet. Example 10: Autologous fat graft Summary
[00145] Fat grafts have recently become more prevalent due to the low low morbidity of the donor site, low complication rate and fast recovery time. In that study, the Depositor examined the role of aspiration and injection pressure in human fat grafts in a nude mouse model. Tumescent liposuction was performed in the laboratory on fresh panniculectomy specimens with a standard 4 mm cannula. The suction pressure was adjusted to -15 inches Hg (-0.5 atmosphere) or -25 inches Hg (-0.83 atmosphere). Liposuction product was centrifuged at 1200G and the fat was injected into the flanks of naked mice with a 16 gauge angiocatheter. Fresh operating room liposuction product was centrifuged at 1200G and the fat was injected into nude mice using low injection pressures and tall. After 4 weeks, the fat lobes were analyzed for weight and histology. Regarding the suction pressure, high versus low suction pressures did not show any apparent differences in weight and histology. Regarding the injection pressure, in a 3 cc syringe, injecting fat at a fast speed (3-5 ml / s) versus a slower speed (0.5-1 cc / s) reaches, in each case, pressures of 2744 mm Hg (3.61 atm) versus 549 mmHg (0.722 atm), (p, 0.001). A low injection pressure provided an improvement of 38% by weight (p <0.001) over the high injection pressure. This was also reflected in histology samples. In conclusion, changes in suction pressures do not affect fat grafts in vivo under the conditions examined. Lobes injected with high pressure, however, did not perform as well as those injected with low pressure. These data indicate that the injection pressure significantly affects the survival of the fat graft. Introduction to the example:
[00146] Autologous fat grafting has become much more prevalent. It has a multitude of cosmetic and reconstructive applications, ranging from breast augmentation to facial haemiatria treatment. Despite its usefulness, fat grafting is limited by unpredictable long-term results. This may require multiple procedures and, as a result, increased risk to the patient. A contributing factor to these disparate results is the wide variety of fat grafting techniques that are currently being used. Improved techniques for autologous fat grafting are required. In this example, the depositor examined the effects of suction and injection pressure on fat grafting.
[00147] The collection of fat grafts can be performed either by liposuction by conventional machine or by liposuction by syringe operated manually. Machine liposuction allows the user to control the suction pressure, which remains constant throughout the collection procedure. In manually operated syringe liposuction, the suction pressure depends on the user. As the syringe fills with liposuction, the vacuum is reduced and the user needs to pull the plunger further back to obtain the same suction pressure. The depositor examined whether there is a difference in graft survival when collection is done with high or low suction pressure.
[00148] The injection pressure can be affected by multiple variables. The depositor found that the injection flow rate is a parameter that can be controlled by the user. Poiseuille's law dictates that as the flow velocity increases, there is a proportional increase in pressure. The depositor examined the effect of increased flow velocity (and pressure) on fat graft survival.
[00149] In this example, the role of aspiration and injection pressure in autologous fat grafts in a small animal model was examined. Materials and methods Suction pressure
[00150] Suction pressure measurements using several syringes (3 ml syringe, 10 ml, 60 ml) were obtained using a manometer (Netech UniMano) (Figure 11). Using a discarded tissue IRB protocol, fresh paniculectomy specimens were obtained from the operating room. Standard tumescent liposuction was performed using pro-machine liposuction (Byron) and a standard 4 mm cannula (Mentor) using -15 inches Hg (-0.5 atmosphere) or -25 inches Hg (-0.83 atmosphere) from suction pressure. Laboratory liposuction product was then centrifuged at 1200 g for 3 minutes in 50 ml of 50 ml conical centrifuge tubes. The fat layer was isolated and a 1 millimeter aliquot was injected into the subcutaneous space of nude mice using a 16-gauge angiocatheter using a 3 ml syringe with a slow injection. After 4 weeks, the animals were sacrificed and the fat lobules were collected for analysis. All animal experimentation was carried out under the protocols described by an approved animal protocol. Injection pressure
[00151] Fresh liposuction product was obtained from the operating room under an IRB protocol of discarded tissue and centrifuged at 1200 G for 3 minutes in 50 ml conical centrifuge tubes. A 1 ml aliquot using a 3 ml syringe was injected into the subcutaneous space of nude mice using a 16-gauge angiocatheter, under an approved animal protocol. The animals were classified into a slow injection group (0.5 - 1.0 ml / s) or a fast injection group (3-5 ml / s). Pressure measurements for fast and slow injections using the same size of angiocatheter and syringe were then obtained using a manometer (Figure 12). After 4 weeks, the animals were sacrificed and the fat lobes were collected. Weight and histology
[00152] The lobes were weighed using a bench scale (Ohaus), after exploding. They were fixed in 10% formalin for 24 hours, processed for paraffin embedding and marked with hematoxylin and eosin. Photographs were taken at WO magnification using a light microscope (Nikon E600). Histology scores were generated by 3 independent, blinded researchers and averaged for each group. The scoring method is based on a previously published scale, which evaluates healthy fat, vacuoles, infiltration product and fibrosis. Each parameter was evaluated based on the following scale: 0 = absence, 1 = minimum presence, 2 = minimum to moderate presence, 3 = moderate presence, 4 = moderate to extensive presence, and 5 = extensive presence. Scores for vacuoles, infiltration product and fibrosis were combined to form a score for total injury. Statistical analysis
[00153] The data are expressed as mean standard error ±. One factor variation analysis was used to compare average weights between experimental groups. The statistical significance was defined by a value of p <0.05. Results Suction pressure
[00154] The maximum suction pressures (negative) obtainable from 3 ml, 20 ml and 60 ml syringes were, in each case, -0.81, -0.92 and 0.96 atmospheres (Figures 13 and 14). In vivo, the average lobed weight aspirated at -15 inches Hg (0.5 atm) versus - 25 inches Hg (0.83 atm) was, in each case, 0.64 +/- 0.03 grams (n = 16) and 0.59 +/- 0.06 grams (n = 16), (p = 0.462). They were not statistically significant (Figure 5). Histological examination showed similar degrees of healthy adipocytes, vacuoles, infiltration product and fibrosis (Figure 16). In addition, the score of these parameters gave a minimal difference (Figure 17) between the group of -15 inches Hg (n = 4_) and the group of -25 inches Hg (n = 3). Injection pressure
[00155] Results of fat pressure injected through a 16-gauge angiocatheter at low speeds (0.5-1.0 ml / s) versus high speeds (3-5 ml./s) were, in each case, 0.72 and 3.61 atmospheres (Fig. 18). In vivo, lobes injected with slow flow rates versus fast flow rates, gave an average weight of, in each case, 0.58 +/- 0.02 grams (n = 32) versus 0.42 +/- 0.02 grams (n = 30) ( p <0.001). This represented an improvement of 38% by weight in slowly injected lobes (Figure 19). Histological appearance also demonstrated healthier adipocytes with fewer vacuoles, an infiltration product and fibrosis (Figure 20). This was also reflected in the determination of scores, which demonstrated a significant increase in healthy adipocyte scores, as well as a significant decrease in injury scores (Figure 21) in slowly injected lobes (n = 17) versus those with rapid injections (n = 13). Exam
[00156] Negative pressure (aspiration) and positive pressure (injection) are two distinct entities. The extremes of pressure ranges obtainable, from zero (that is, perfect vacuum or outer space) to infinity (theoretical). At or near sea level, the atmospheric pressure is equal to 1 atmosphere. The negative (or positive) pressure obtained in a syringe is the difference between inside and outside the syringe. Therefore, when the pressure inside a syringe is reduced to 0.75 atmospheres and the external pressure is 1 atmosphere (sea level), then the negative pressure obtained is -0.25 atmosphere. Since the minimum obtainable absolute pressure is zero, then the maximum negative pressure obtainable at sea level is -1 atmosphere. This does not apply to positive pressure, because there is no theoretical maximum pressure. Therefore, changes in pressure can be much greater when fat is injected.
[00157] A second applicable principle is that a liposuction by syringe operated manually depends mainly on Boyle's law, which states: Pressurei x Volume 1 = Pressure2X Volume 2
[00158] Therefore, the pressure change is dependent on the relationship between the initial volume and the final volume. When the syringe plunger starts at the zero mark, then the initial volume is equal to the volume inside the suction cannula (often less than one ml). This concept is important when measuring suction pressures with a manometer, because the initial volume needs to be similar. Inserting any tube between the pressure gauge and the syringe increases the initial volume sufficiently to produce inaccurate results. The measured pressures obtained with several syringes (Figures 13 and 14) followed this principle and changed essentially only in relation to the change in volume. Differences in pressure obtained were minimal and all approached - 1 atmosphere, but did not exceed the same.
[00159] Traditionally manually operated syringe liposuction is performed by withdrawing approximately a2 ml in a 10 ml syringe. The data presented in the present show that this is approximately equivalent to -15 to - 20 inches Hg adjusted on a liposuction machine (Figure 13).
[00160] In the animal model, fat collected at - 15 inches Hg, a typical manually operated syringe liposuction pressure and 25 inches Hg, a typical machine liposuction pressure, did not show a difference in either weight or histology (Figures 15, 16 and 17). This is clinically relevant, at least because the high pressure machine liposuction can be used without fear of affecting the viability of the fat graft. This can often be an easier and more efficient method of collecting fat grafts.
[00161] As previously mentioned, the injection pressure can be affected by many variables, including flow rate. Injecting fat through a 16 gauge angiocatheter at a rapid flow rate (3-5 ml / s) produced a pressure five times that of the slow injection (0.5-1.0 ml / s). In our animal model, slowly injected fat grafts improved 38% by weight, when compared to those injected with a fast flow rate. These fat grafts also looked healthier in histology, with less vacuoles, fibrosis and infiltration product. Table 1 shows injection rates at different flow rates and catheter sizes. Table 1: Injection speeds through several catheters


[00162] Without wishing to be limited in theory, a possible factor, which contributes to the observation and that the injection pressure affects the fat graft overexperience, while the suction pressure does not, is that the pressure has a direct traumatic effect on adipocytes, which ultimately causes cell death and graft reabsorption. This can be explained by the fact that the maximum negative pressure obtained is - 1 atmosphere, while the maximum obtainable injection pressure is much higher. In our experiments, the average pressure of a rapid injection was 3.61 atmospheres. Therefore, it is possible to apply a much higher pressure on grafts during injection than during suction / aspiration. By minimizing the injection pressure, and resulting shear loading speed, trauma to fat grafts can be minimized, thereby improving the survival of fat grafts. Conclusions
[00163] Changes in suction / aspiration pressures do not affect fat grafts in vivo under the conditions examined. Lobes injected at low pressure, however, produced an improvement of 38% by weight and had an improved appearance in histology. These data indicate that an injection pressure significantly affects the survival of fat grafts.
[00164] Having thus described several aspects of at least one modality of this invention, it should be understood that several changes, modifications and improvements are easily apparent to those skilled in the art. These changes, modifications and improvements are intended to be part of this description and are intended to be within the spirit and scope of the invention. Consequently, the above description and drawings are by way of example only and the invention is described in detail by the following claims.
[00165] Certain embodiments of the present invention provided for in the present are described mainly in terms of treatment and purification of adipose tissue, to obtain samples of viable adipose cells for transplantation. However, these modalities can be used to treat or process other biological materials, such as other tissues or cells (for example, basal cells).
[00166] The use of ordinal terms, such as "first", "second", "third" etc. in claims to modify an element of king-vindication, it does not in itself indicate any priority, precedence or order of one element of claim over another or the temporal order in which acts of a method are performed, but are used only as a mark to distinguish a claim element with a given name from another element with the same name (except for the use of the term ordinal) to distinguish claim elements.

权利要求:
Claims (29)
[0001]
1. Apparatus characterized by comprising a chamber with an entrance and a first exit; a retention matrix within the chamber configured and arranged to contact a biological material, when present in the chamber, so that a fraction of the biological material is retained in the retention matrix, where the retention matrix comprises at least one selected from the group of a lipophilic matrix that retains lipids from biological material and a hydrophilic matrix that retains water from biological material; and a filter within the chamber that is configured and arranged to contact biological material, when present in the chamber, so that a residual fraction of the biological material passes through the filter; wherein the inlet is fluidly connected to the first outlet along a first flow path that does not pass through the filter.
[0002]
2. Apparatus, according to claim 1, is characterized by the fact that the first outlet is fluidly connected to an appropriately sized cannula or catheter.
[0003]
3. Apparatus, according to claim 2, is characterized by the fact that the cannula or catheter of appropriate size is 12 gauge, 14 gauge, 15 gauge, 16 gauge, 17 gauge or 18 gauge.
[0004]
4. Apparatus, according to claim 1, is characterized by the fact that the chamber is configured and arranged to contain 1 ml to 1 L of biological material.
[0005]
5. Apparatus, according to claim 1, is characterized by the fact that biological material is present in the chamber.
[0006]
6. Apparatus, according to claim 1, is characterized by the fact that it also comprises biological material, in which the biological material comprises adipose tissue or a component thereof.
[0007]
7. Apparatus, according to claim 6, is characterized by the fact that the adipose tissue or a component thereof comprises adipocytes, adipogenic cells, mesenchymal cells or stem cells.
[0008]
8. Apparatus according to claim 1, characterized by the fact that it further comprises a membrane stabilizing agent (MSA) in the chamber.
[0009]
9. Apparatus according to claim 8, characterized by the fact that the MSA is a tri-block copolymer comprising a tri-block copolymer of the form: polyethylene glycol-polypropylene glycol-polyethylene glycol.
[0010]
10. Apparatus according to claim 9, characterized by the fact that MSA is poloxamer P188.
[0011]
11. Apparatus according to claim 8, characterized by the fact that the retention matrix retains at least one selected from the group of water, lipids, metals, membrane stabilizing agent and blood cells within the retention matrix .
[0012]
12. Apparatus according to claim 1, characterized by the fact that it further comprises a pressure generating device that controls a pressure within the chamber.
[0013]
13. Apparatus according to claim 12, characterized by the fact that the pressure generating device is a pump.
[0014]
14. Apparatus, according to claim 13, characterized by the fact that it also comprises a controller configured and arranged to generate control signals that activate the pump to generate a positive pressure in the chamber and a pressure sensor fluidly connected to the chamber and with an electrical output connected to a controller input, the pressure sensor that provides an electrical signal to the controller indicating a pressure detected in the chamber, where the controller transmits control signals to the pump based on the detected pressure.
[0015]
15. Apparatus according to claim 12, characterized by the fact that the pressure generating device is configured and arranged to generate a positive pressure within the chamber that is equal to or less than a predetermined threshold of about 6 atm, the positive pressure being sufficient to cause the biological material, if present in the chamber, to discharge through the outlet, where the predetermined threshold of about 6 atm is a pressure above which the biological material has a relatively low viability as a graft of tissue after discharge from the outlet to a graft site on a subject.
[0016]
16. Apparatus according to claim 15, characterized by the fact that the positive pressure is maintained so that the speed of the biological material discharged from the outlet is up to about 265 cm / s.
[0017]
17. Apparatus according to claim 15, characterized by the fact that the positive pressure is maintained so that the flow rate of the biological material being discharged from the outlet is less than 3 ml / s.
[0018]
18. Apparatus according to claim 15, characterized by the fact that the positive pressure is about 4 atm.
[0019]
19. Apparatus according to claim 1, characterized by the fact that the retention matrix includes the hydrophilic matrix and the lipophilic matrix.
[0020]
20. Apparatus according to claim 19, characterized by the fact that the hydrophilic matrix is positioned downstream of the lipophilic matrix within the chamber.
[0021]
21. Apparatus according to claim 1, characterized by the fact that the first outlet of the chamber is configured and arranged to allow the discharge of a portion of the biological material that does not pass through the filter.
[0022]
22. Apparatus according to claim 1, characterized by the fact that the chamber has a second waste outlet, in which the inlet is connected fluidly to the second waste outlet along a second flow path through the filter and of the retention matrix.
[0023]
23. Apparatus according to claim 22, characterized by the fact that the filter is disposed between the second waste outlet and the retaining matrix.
[0024]
24. Apparatus according to claim 22, characterized by the fact that the retention matrix and filter are configured and arranged so that the residual fraction of biological material flows through the filter and the retention matrix to the second outlet waste.
[0025]
25. Method, characterized by the fact that it comprises: obtaining the apparatus of claim 12; loading biological material into the device chamber; and causing the pressure generating device to generate a positive pressure within the chamber that is at or below a predetermined threshold of about 6 atm to cause the biological material to discharge through the outlet.
[0026]
26. Method, according to claim 25, characterized by the fact that the biological material comprises adipose tissue and the method further comprises positioning the outlet, so that the biological material is discharged at a subject's graft site.
[0027]
27. Method, according to claim 26, characterized by the fact that the biological material is obtained from the subject.
[0028]
28. Method, according to claim 27, characterized by the fact that the biological material is obtained by extraction of adipose tissue from the abdomen, thigh, flank region or gluteal region of the subject or by extraction of liposuction that comprises this adipose tissue of the subject.
[0029]
29. Method according to claim 26, characterized by the fact that biological material is obtained from the subject and processed before loading into the chamber to remove at least partially oil, blood, tumescent fluid, stromal cells, extracellular material or cellular debris from biological material.

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公开号 | 公开日

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CN108504539A|2018-09-07|

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JP6072684B2|2017-02-01|

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KR101809301B1|2018-01-18|

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JP2013538081A|2013-10-10|

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EP2601299B1|2019-04-24|

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EP3530741B1|2021-04-07|

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KR20140023247A|2014-02-26|

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EP2601299A4|2014-03-19|

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CN103153376B|2018-04-20|

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BR112013002859A2|2016-05-31|

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CN108504539B|2021-11-02|

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WO2012019103A2|2012-02-09|

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US20130158515A1|2013-06-20|

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WO2021110282A1|2019-12-04|2021-06-10|HYDRA S.r.l.|Device for the fragmentation of tissues within a sealed sterile environment with an aseptic procedure and method thereof|

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CN113116414A|2021-04-22|2021-07-16|任学会|Full closed system for autologous fat filling transplantation|

法律状态:
2018-12-26| B06F| Objections, documents and/or translations needed after an examination request according [chapter 6.6 patent gazette]|

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2019-09-17| B06U| Preliminary requirement: requests with searches performed by other patent offices: procedure suspended [chapter 6.21 patent gazette]|

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2020-05-19| B09A| Decision: intention to grant [chapter 9.1 patent gazette]|

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2020-10-06| B16A| Patent or certificate of addition of invention granted [chapter 16.1 patent gazette]|Free format text: PRAZO DE VALIDADE: 20 (VINTE) ANOS CONTADOS A PARTIR DE 05/08/2011, OBSERVADAS AS CONDICOES LEGAIS. |

优先权:

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申请号 | 申请日 | 专利标题

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US37140010P| true| 2010-08-06|2010-08-06|

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US61/371,400|2010-08-06|

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PCT/US2011/046752|WO2012019103A2|2010-08-06|2011-08-05|System and apparatus for cell treatment|

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