WO2019067942A1 - Implants bioactifs et procédés de fabrication - Google Patents

Implants bioactifs et procédés de fabrication Download PDF

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Publication number
WO2019067942A1
WO2019067942A1 PCT/US2018/053495 US2018053495W WO2019067942A1 WO 2019067942 A1 WO2019067942 A1 WO 2019067942A1 US 2018053495 W US2018053495 W US 2018053495W WO 2019067942 A1 WO2019067942 A1 WO 2019067942A1
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Prior art keywords
medical implant
subject
medical
implant
tissue
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PCT/US2018/053495
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English (en)
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Amit Prakash Govil
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Amit Prakash Govil
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Publication of WO2019067942A1 publication Critical patent/WO2019067942A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/39—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin, cold insoluble globulin [CIG]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/35—Fat tissue; Adipocytes; Stromal cells; Connective tissues
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/36—Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/18—Growth factors; Growth regulators
    • A61K38/1858—Platelet-derived growth factor [PDGF]
    • A61K38/1866—Vascular endothelial growth factor [VEGF]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
    • A61L27/3608—Bone, e.g. demineralised bone matrix [DBM], bone powder
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
    • A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • A61L27/3804—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
    • A61L27/3834—Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412—Tissue-regenerating or healing or proliferative agents
    • A61L2300/414—Growth factors

Definitions

  • BACKGROUND Correction of subcutaneous soft tissue defects in humans is a significant problem that faces the field of medicine, especially that of plastic surgery.
  • implants which aim to correct such defects, have limited capacity to interact with the environment in which they are implanted. This limited capacity to interact with the host tissue precludes such implants from reaching their full potential, and in many cases, can prevent successful outcomes for such implants.
  • the implantation of existing implants can be invasive, causing undue trauma at the site of implantation, and the implantation of implants that can maintain a desired shape after implantation, while staying functional, is lacking.
  • bioactive implants suffer from poor flowability through traditional applicators or injectors, impeding successful implantation of such. As such, there exists a need for improved bioactive implants, as well as methods of making bioactive implants in addition to methods for delivery of medical implants.
  • Bioactive implants can comprise collagen, hyaluronan, protein, amino acid, peptide, or glycosaminoglycan, individually or in combination.
  • Medical implants can further comprise biocompatible material and/or growth factors.
  • Medical implants can include a plurality of cells from one or more sources. Cells or growth factors of medical implants as described herein can be each independently derived from adipose, bone marrow, blood, amniotic fluid, or from another matrix such as intestine, bladder, placental, or other soft tissue. Medical implants can be acellular and/or delipidized.
  • Medical implants as described herein can comprise decontaminated adipose, dermis, or fascia, individually or in combination. Medical implants can be derived from allograft, autograft, xenograft, or synthetic sources. The source of cells or growth factors of medical implants as described herein can be a different source than the other components of the medical implant.
  • Described herein are methods of treatment.
  • a method of treating pain in a subject can comprise delivering medical implants as described herein to one or more vertebral discs in the spine of a subject in need thereof in a therapeutic amount.
  • Methods of treatment and/or treating pain can comprise delivering medical implants as described herein to a joint of a subject in need thereof in a therapeutic amount.
  • Methods of treatment as described herein can comprise delivering medical implants as described herein to a subject in need thereof in a therapeutic amount.
  • methods of treatment can be methods of treating soft-tissue defects.
  • Methods of treating soft-tissue defects can comprise delivering medical implants as described herein to a subject in need thereof in a therapeutic amount.
  • Described herein are methods of providing support or cushioning to treat soft-tissue defects in a subject. Such methods can comprise delivering medical implants as described herein to a subject in need thereof in a therapeutic amount.
  • Methods of increasing growth factor binding can comprise providing a bone composition in a solution; adding an acid to the solution, thereby exposing the bone composition to an acid to demineralize the bone composition; and adding a buffer or base to raise the pH of the solution from a first level to a second level, wherein the second level is more basic than the first level, after at least partial demineralization of the bone.
  • Methods to increase growth factor binding can also comprise isolating the bone composition, mineral that has precipitated of solution, or both.
  • the extracellular matrix of the bone and/or bone composition can be removed prior to adding an acid to the solution.
  • kits containing medical implants as described herein can comprise of medical implants as described herein and a delivery device (such as an applicator; also referred to as an applicator).
  • a delivery device such as an applicator; also referred to as an applicator.
  • the medical implant is loaded in the delivery device in the kit and comes ready for a user's use.
  • Medical implants as described herein can be medical implants prepared according to any of the methods as described herein, individually or in combination.
  • Such methods comprise providing a scaffold in solution; combining the scaffold in solution with growth factors or agents; and raising the pH of the solution from a first level to a second level, wherein the second level is more basic than the first, by adding a base or buffer to the solution.
  • Such methods can further comprise isolating the scaffold.
  • the scaffold can be derived from allograft or xenograft tissue sources or created synthetically.
  • the scaffold can be comprised of comprised of collagen, hyaluronan, or other protein, amino acid, GAG, and/or biocompatible material.
  • Growth factors or agents can comprise VEGF, bFGF, aFGF,TGFB-1 , PDGF, IGF, or any other growth factor or tissue growth stimulative agent or differentiation agent, individually or in combination.
  • Growth factors or agents comprise VEGF, bFGF, aFGF.TGFB- 1 , PDGF, or IGF, individually or in combination.
  • Methods for providing shape memory for a medical implant can comprise providing the medical implant; and providing energy to the medical implant.
  • the medical implant can be frozen before providing the energy.
  • the medical implant can be hydrated before providing the energy.
  • One or more additives can be added to the medical implant, before providing the energy or after providing the energy.
  • Additives can be one or more of a vasodilator, a growth factor, or an amino acid.
  • Medical implants can be placed in a mold of a desired shape before providing the energy.
  • the mold can be constructed of a polymer or other material that can be confirmed to a shape and then becomes rigid after conforming to the shape.
  • the mold can be configured to receive the medical implant in a desired configuration according to the shape of the destination to which the implant will be implanted.
  • Medical implants can be freeze dried before or after providing the energy.
  • Medical implants can be sterilizing after providing the energy.
  • Medical implants can be terminally sterilized after the energy is provided.
  • Medical implants as described herein can be three-dimensional medical implants. Medical implants as described herein can be derived from an allograft. Medical implants as described herein can be primarily comprised of collagen. Medical implants as described herein can be micronized soft tissue. Medical implants as described herein can be a particulated medical implant.
  • FIG. 1 is an embodiment of an applicator 100 according to the present disclosure.
  • FIG. 2 shows a portion of an embodiment of a needle 101 of an applicator 100 according to the present disclosure, from a side of the applicator 100 different than what is shown in FIG. 1.
  • FIG. 3 is another embodiment of an applicator according to the present disclosure.
  • FIG. 4 is an embodiment 200 according to the present disclosure.
  • FIG. 5 is an embodiment 300 according to the present disclosure.
  • FIG. 6 is a flow chart illustrating an embodiment of the method 1000.
  • FIG. 7 is a flow chart illustrating an embodiment of the method 2000.
  • FIG. 8 is a flow chart illustrating an embodiment of the method 3000.
  • FIG. 9 is a flow chart illustrating an embodiment of the method 4000.
  • FIG. 10 is a flow chart illustrating an embodiment of the method 5000.
  • FIG. 1 1 is a graph depicting the modulation of flow characteristics in terms of mix energy.
  • FIG. 12 is a graph of volume retention versus non-protein/GAG mass according to an embodiment of the present disclosure.
  • FIGs. 13A and 13B are before (FIG. 13A) and after (FIG. 13B) thermal images taken to visualize temperature differences created by subcutaneous fat insulation.
  • FIG. 13A was taken of the subject before application and
  • FIG. 13B was taken of the subject about a week later. Notice the change on one side of the body.
  • FIG. 14 is an example of a crosslinked shape-formed medical implant according to the present disclosure.
  • Embodiments of the present disclosure will employ, unless otherwise indicated, techniques of molecular biology, physiology, modern surgical techniques, microbiology, organic chemistry, biochemistry, and the like, which are within the skill of the art. Such techniques are explained fully in the literature.
  • effective amount is an amount sufficient to effect beneficial or desired results.
  • An effective amount can be administered in one or more administrations, applications, or dosages.
  • therapeutic refers to treating or curing a disease or condition.
  • preventative refers to hindering or stopping a disease or condition before it occurs or while the disease or condition is still in the sub-clinical phase.
  • concentration used in reference to an amount of a molecule, compound, or composition, including, but not limited to, a chemical compound, polynucleotide, peptide, polypeptide, protein, antibody, or fragments thereof, that indicates that the sample is distinguishable from its naturally occurring counterpart in that the concentration or number of molecules per volume is greater than that of its naturally occurring counterpart.
  • isolated means separated from constituents, cellular and otherwise, with which the polynucleotide, peptide, polypeptide, protein, antibody, or fragments thereof, are normally associated in nature.
  • a non-naturally occurring polynucleotide, peptide, polypeptide, protein, antibody, or fragments thereof, does not require “isolation” to distinguish it from its naturally occurring counterpart.
  • diluted used in reference to an amount of a molecule, compound, or composition including but not limited to, a chemical compound, polynucleotide, peptide, polypeptide, protein, antibody, or fragments thereof, that indicates that the sample is distinguishable from its naturally occurring counterpart in that the concentration or number of molecules per volume is less than that of its naturally occurring counterpart.
  • subject refers to a vertebrate, preferably a mammal, more preferably a human. Mammals include, but are not limited to, murines, simians, humans, farm animals, sport animals, and pets.
  • the term “pet” includes a dog, cat, guinea pig, mouse, rat, rabbit, ferret, and the like.
  • farm animal includes a horse, sheep, goat, chicken, pig, cow, donkey, llama, alpaca, turkey, and the like.
  • biocompatible or “biocompatibility” refers to the ability of a material to be used by a patient without eliciting an adverse or otherwise inappropriate host response in the patient to the material or a derivative thereof, such as a metabolite, as compared to the host response in a normal or control patient.
  • progeny As used herein, “cell,” “cell line,” and “cell culture” include progeny. It is also understood that all progeny may not be precisely identical in DNA content, due to deliberate or inadvertent mutations. Variant progeny that have the same function or biological property, as screened for in the originally transformed cell, are included.
  • binding refers to binding which occurs between such paired species as enzyme/substrate, receptor/agonist, antibody/antigen, and lectin/carbohydrate which may be mediated by covalent or non-covalent interactions or a combination of covalent and non-covalent interactions.
  • the binding which occurs is typically electrostatic, hydrogen- bonding, or the result of lipophilic interactions. Accordingly, “specific binding” occurs between a paired species where there is interaction between the two which produces a bound complex having the characteristics of an antibody/antigen or enzyme/substrate interaction.
  • the specific binding is characterized by the binding of one member of a pair to a particular species and to no other species within the family of compounds to which the corresponding member of the binding member belongs.
  • an antibody preferably binds to a single epitope and to no other epitope within the family of proteins.
  • control is an alternative subject or sample used in an experiment for comparison purposes and included to minimize or distinguish the effect of variables other than an independent variable.
  • positive control refers to a “control” that is designed to produce the desired result, provided that all reagents are functioning properly and that the experiment is properly conducted.
  • negative control refers to a “control” that is designed to produce no effect or result, provided that all reagents are functioning properly and that the experiment is properly conducted.
  • Other terms that are interchangeable with “negative control” include “sham,” “placebo,” and “mock.”
  • culturing refers to maintaining cells under conditions in which they can proliferate and avoid senescence as a group of cells. “Culturing” can also include conditions in which the cells also or alternatively differentiate.
  • “synergistic effect,” “synergism,” or “synergy” refers to an effect arising between two or more molecules, compounds, substances, factors, or compositions that is greater than or different from the sum of their individual effects.
  • additive effect refers to an effect arising between two or more molecules, compounds, substances, factors, or compositions that is equal to or the same as the sum of their individual effects.
  • autologous refers to being derived from the same subject that is the recipient.
  • graft refers to a graft that is derived from one member of a species and grafted in a genetically dissimilar member of the same species.
  • xenograft or “xenogeneic” refers to a substance or graft that is derived from one member of a species and grafted or used in a member of a different species.
  • autograft refers to a graft that is derived from a subject and grafted into the same subject from which the graft was derived.
  • allogeneic refers to involving, derived from, or being individuals of the same species that are sufficiently genetically different so as to interact with one another antigenicaly.
  • “syngeneic” refers to subjects or donors that are genetically similar enough so as to be immunologically compatible to allow for transplantation, grafting, or implantation.
  • implant or "graft,” as used interchangeably herein, refers to cells, tissues, or other compounds, including metals and plastics, that are inserted into the body of a subject.
  • filler refers to a substance used to fill a cavity or depression.
  • the filler can fill the depression such that it is level with the surrounding area or that the cavity is filled, such that the depth of the depression or volume of the cavity is decreased, or such that the area that was the depression is now raised relative to the areas immediately surrounding the depression.
  • immunological refers to the ability of a substance, compound, molecule, and the like (referred to as an "antigen") to provoke an immune response in a subject.
  • exogenous refers to a compound, substance, or molecule coming from outside a subject or donor, including their cells and tissues.
  • endogenous refers to a compound, substance, or molecule originating from within a subject or donor, including their cells or tissues.
  • bioactive refers to the ability or characteristic of a material, compound, molecule, or other particle that interacts with or causes an effect on any cell, tissue and/or other biological pathway in a subject.
  • bioactive factor refers to a compound, molecule, or other particle that interacts with or causes an effect on any cell, tissue, and/or other biological pathway in a subject.
  • physiological solution refers to a solution that is about isotonic with tissue fluids, blood, or cells.
  • donor refers to a subject from which cells or tissues are derived.
  • slurry refers to the resultant product from any of the methods described herein. Accordingly, the slurry can be in any form resulting from the processing described herein, including but not limited to, dehydrated slurry or tissue, paste, powder, solution, gel, putty, particulate and the like.
  • extra cellular matrix refers to the non-cellular component surrounding cells that provides support functions to the cell including structural, biochemical, and biophysical support, including but not limited to, providing nutrients, scaffolding for structural support, and sending or responding to biological cues for cellular processes such as growth, differentiation, and homeostasis.
  • complete extracellular matrix refers to extracellular matrix that has all components (proteins, peptides, proteoglycans, and the like) present and may or may not include other cells that are embedded in the extra cellular matrix.
  • decellularized extracellular matrix refers to complete extracellular matrix that has been processed to remove any cells embedded within the extracellular matrix.
  • extracellular matrix component refers to a particular component.
  • an extracellular matrix comportment can be a a specific class of comments (e.g. proteoglycans) or individual component (e.g. collagen I) that is separated or isolated from the other extracellular components. These components can be made synthetically.
  • hydrogel refers to a network of hydrophilic polymer chains that are dispersed in water.
  • Hydrophilic polymer chains dispersed in water that are found as a colloidal gel.
  • self-assembling peptides refer to peptides which undergo spontaneous assembly into ordered nanostructures.
  • Self-assembling peptides include di- peptides, lego peptides, surfactant peptides, molecular paint or carpet peptides, and cyclic peptides.
  • administering refers to an administration that is oral, topical, intravenous, subcutaneous, transcutaneous, transdermal, intramuscular, intra-joint, parenteral, intra-arteriole, intradermal, intraventricular, intracranial, intraperitoneal, intralesional, intranasal, rectal, vaginal, by inhalation or via an implanted reservoir.
  • parenteral includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injections or infusion techniques.
  • effective amount refers to an effective amount of medical implants as described herein to reduce the appearance of subcutaneous soft-tissue defects, to bolster the support of endogenous soft-tissue in the body which supports joints (for example, ankle joints, knee joints, vertebral discs, and the like, or to aid in the cushioning provided by soft- tissue at joints or on pads of the hands and/or feet.
  • primarily means that a part of the whole is present in an amount greater than all other parts of the whole, greater than all other parts individually or in combination.
  • bioactive implants also referred to herein as medical implants
  • methods of making methods of delivery, and kits comprising bioactive implants.
  • medical implants can comprise collagen, hyaluronan, protein, amino acid, peptide, and glycosaminoglycan.
  • the medical implants are primarily collagen-based and derived from allograft tissue.
  • proteins and peptides are proteins and peptides derived from allograft tissue.
  • medical implants can further comprise biocompatible material.
  • biocompatible material can be material that can product medical implants during processing and/or storage.
  • the biocompatible material is one or more of glycerol, propylene glycol, sugars, and preservatives, individually or in combination.
  • medical implants can further comprise growth factors.
  • medical implants can comprise a plurality of cells from one or more sources.
  • the cell or growth factors of the present disclosure can each independently be derived from adipose, bone marrow, blood, muscle, amniotic fluid, or from another matrix such as intestine, bladder, placental, or other soft tissue.
  • Sources of such can be allograft, autograft, xenograft, or synthetic sources.
  • medical implants are acellular.
  • medical implants can be delipidized. Lipids can be removed from medical implants according to well-known methods as known by skilled artisans.
  • medical implants as described herein can comprise decontaminated adipose, dermis, or fascia, individually or in combination.
  • Medical implants as disclosed herein can be derived from allograft, autograft, xenograft, or synthetic sources.
  • a method of treating pain in a subject can comprise delivering the medical implant of any one of claims 1 to 9 to a vertebral disc in the spine of a subject in need thereof in a therapeutic amount.
  • a subject in need thereof can be a human subject experiencing pain.
  • a subject in need thereof can be a human subject experiencing joint (knee, ankle, wrist, any joint of the human body).
  • a therapeutic amount can be about 0.1 cc to about 5cc.
  • a therapeutic amount can be about 0.5cc to about 4.5cc.
  • a therapeutic amount can be about 1cc to about 4cc.
  • a therapeutic amount can be about 1.5cc to about 3.5cc.
  • a therapeutic amount can be about 2cc to about 3cc.
  • a therapeutic amount can be about 2.5cc.
  • a method of treating pain in a subject can comprise delivering a medical implant as described herein to a knee of a subject in need thereof in a therapeutic amount.
  • a subject in need thereof can be a human subject experiencing knee pain.
  • a therapeutic amount can be 0.1 cc to 5cc.
  • a therapeutic amount can be about 0.5cc to about 4.5cc.
  • a therapeutic amount can be about 1cc to about 4cc.
  • a therapeutic amount can be about 1.5cc to about 3.5cc.
  • a therapeutic amount can be about 2cc to about 3cc.
  • a therapeutic amount can be about 2.5cc.
  • a method of treating pain in a subject can comprise delivering a medical implant as described herein to one or more vertebral discs of a subject in need thereof in a therapeutic amount.
  • a subject in need thereof can be a human subject experiencing spinal and/or back pain, or other peripheral nerve pain as a result of nerve compression by vertebrae of the spine.
  • a therapeutic amount can be 0.1 cc to 5cc.
  • a therapeutic amount can be about 0.5cc to about 4.5cc.
  • a therapeutic amount can be about 1 cc to about 4cc.
  • a therapeutic amount can be about 1.5cc to about 3.5cc.
  • a therapeutic amount can be about 2cc to about 3cc.
  • a therapeutic amount can be about 2.5cc.
  • Methods as described herein can comprise delivering the medical implant to a subject in need thereof in a therapeutic amount.
  • Methods as described herein can comprise delivering the medical implant of any one of claims 1 to 9 to a subject in need thereof in a therapeutic amount.
  • Methods of increasing growth factor binding as described herein can comprise providing a bone composition in a solution; adding an acid to the solution, thereby exposing the bone composition to an acid to demineralize the bone composition; adding a buffer or base to raise the pH of the solution in the presence of one or more growth factors from a first level to a second level, wherein the second level is more basic than the first level, after at least partial demineralization of the bone.
  • the bone composition can then be isolated, as can the mineral that has precipitated of solution (for example calcium), or both.
  • the bone extracellular matrix is removed prior to raising the pH in order to precipitate the calcium from the bone.
  • Buffers or bases as described herein can raise the pH of a composition or a solution from a first level to a second level, wherein the second level is more basic than the first level.
  • buffers or bases as described herein can comprise sodium hydroxide, water, sodium bicarbonate, phosphate buffer, sodium chloride, or calcium salt.
  • Bone compositions as described herein can comprise mineralized bone, cancellous bone, or both.
  • the first pH level is an acidic pH level.
  • medical implants as described herein are medical implants according to or created by any of the methods as described herein, individually or in combination.
  • Kits as described herein can comprise medical implants are described according to any of the methods as described herein, individually or in combination, and an applicator. In certain aspects, the medical implant of the kit is pre-loaded into the applicator.
  • Applicators that can implant medical implants as described herein.
  • Applicators as described herein can be flexible applicators that can allow for accurate delivery of a fluid, gel, or scaffold.
  • the applicator may contain a flexible tip, plunger/wire, crimped tube, and the like.
  • Non-implantable portions of the applicator are configured to be removed after placement of the implant in the desired location of the subject.
  • Embodiments of applicators as described herein can be used to implant medical implants and can be used to fill defects in skin, adipose, muscle, vasculature, or other soft tissue void.
  • FIG. 1 shows an embodiment of an applicator 100 according to the present disclosure.
  • the applicator 100 comprises a needle 101 (which can be a trocar needle and can have an opening along a longitudinal axis A in addition to the end B, as shown in FIG. 2) attached to a thread 105 via a mechanical joint 103 (the mechanical joint 103 can be a mechanical crimp joint).
  • the thead 105 can be dehydrated and can be different sizes or lengths, but can be about 0.1 to about 1.0 mm in diameter or greater.
  • the needle 101 can be a trocar needle and can be about 1 gauge to about 33 gauge on the Stubbs scale.
  • the needle 101 is a 27 gauge trocar (7.5 cm).
  • the applicator 100 can be inserted subcutaneously into a dermal layer of a subject that contains soft-tissue defects, which are to be corrected.
  • the applicator 100 can then be loaded with the medical implant, or the applicator 100 can be preloaded with the medical implant to be implanted.
  • the medical implant can be implanted with the applicator 100 by the addition of pressure (by way of a mechanical syringe or stylet coupled to the applicator 100), and the applicator 100 can then be removed from the dermal layer of the subject, leaving behind the medical implant.
  • the epidermal layer of a subject can have a defect, such as a scar or a ripple, and the applicator 100 can be inserted subcutaneously below the defect.
  • the applicator 100 can then implant the medical implant subcutaneously below the defect, and the applicator can then be removed from the subject.
  • FIG. 3 is a photograph showing another embodiment of an applicator as described herein.
  • an applicator as described herein can comprise a syringe and a stylet for delivering the medical implant into the subject.
  • an applicator as described herein can comprise a hollow tube that may be curved to match the precise anatomy of a subject in a region of interest where there is a soft-tissue defect.
  • the hollow tube can have a funnel at an end that is not inserted into the subject, and an outer ring at the opposite end that is to be inserted into the subject.
  • a stylet can then be used to precisely deliver medical implants as described herein into the subject with the applicator.
  • FIG. 4 shows an embodiment according to the present disclosure 200.
  • a medical implant is a medical implant that is cut and/or otherwise formed into a tube 201 (a three-dimensional structure with a longitudinal axis of greater dimension than the diameter and/or circumference that is substantially circular when viewed in a direction along the longitudinal axis).
  • a needle or wire 203 (or applicator 100) can be inserted into the allograft medical implant tube 201 along the longitudinal axis C or in a hole of the allograft medical implant tube 201 along the longitudinal axis C to provide rigidity.
  • the allograft tube 201 can then be implanted into the location of interest in a subject, and the needle/wire 203 can be removed, leaving the medical implant in place in the subject while the need/wire 203 or applicator 100 is removed from the subject.
  • the needle or wire 203 or applicator 100 can be packaged in a kit with the allograft tube 203, or a kit can comprise the needle or wire 203 or applicator 100 pre-loaded in the allograft medical implant tube 201.
  • FIG. 5 shows another embodiment according to the present disclosure.
  • a medical implant comprises allograft particles/pieces 303 that are preloaded into the hollow shaft of a needle 301 , the needle 301 comprising an implantation end E and a non-implanted end F.
  • the hollow shaft of the needle 301 is also configured to receive a stylet 305 on an end that is not implanted into the subject.
  • the implantation end E of the needle is inserted into a subject and positioned to where the medical implant is intended to be implanted in the subject.
  • Described herein is method of enriching bound growth factors in a medical implant, comprising: providing a scaffold in solution; combining the scaffold in solution with growth factors or agents; raising the pH of the solution from a first level to a second level, wherein the second level is more basic than the first, by adding a base or buffer to the solution. After the pH is made more basic, the scaffold can be isolated.
  • the scaffold can be derived from allograft or xenograft tissue sources or created synthetically.
  • the scaffold can be comprised of comprised of collagen, hyaluronan, or other protein, amino acid, glycosaminoglycan (GAG), and/or biocompatible material (for example, calcium, salt, glycerol, or other storage agents or biological preservatives known in the art).
  • FIG. 6 is a flow chart illustrating an embodiment of the method 1000.
  • a bone composition is provided in a solution 1010.
  • the bone composition can comprise cortical bone, cancellous bone, or both.
  • An acid can then be added to the solution 1020, followed by the addition of a base or buffer to the solution 1030 to raise the pH (make it more basic).
  • the buffer or base can comprise one or more of sodium hydroxide, water, sodium bicarbonate, phosphate buffer, sodium chloride, or calcium salt.
  • the bone composition can then be isolated after the pH is raised. Acids used may be weak or strong. Acids may include, but not limited to hydrochloric or phosphoric, sulphuric, glycolic, acetic, paracetic, citric, ascorbic, and the like.
  • the solution can further comprise one or more growth factors.
  • the method can further comprise adding one or more growth factors at any stage of the method.
  • Growth factors can comprise vasoendothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), acidic fibroblast growth factor (aFGF), transforming growth factor beta-1 (TGFB-1), platelet-derived growth factor (PDGF), insulin-like growth factor IGF, or any other growth factor or tissue growth stimulative agent or differentiation agent, individually or in combination, or other growth factors as described herein.
  • VEGF vasoendothelial growth factor
  • bFGF basic fibroblast growth factor
  • aFGF acidic fibroblast growth factor
  • TGFB-1 transforming growth factor beta-1
  • PDGF platelet-derived growth factor
  • IGF insulin-like growth factor
  • FIG. 7 is a flow chart illustrating an embodiment of the method 2000.
  • a bone composition is provided in a solution 2010.
  • the bone composition can comprise cortical bone, cancellous bone, or both.
  • An acid can then be added to the solution 2015, and the extracellular matrix (ECM) is then removed from the bone composition 2020.
  • ECM extracellular matrix
  • a base or buffer to the solution 2030 to raise the pH (make it more basic).
  • the buffer or base can comprise one or more of sodium hydroxide, water, sodium bicarbonate, phosphate buffer, sodium chloride, or calcium salt.
  • the bone composition can then be isolated after the pH is raised. Acids used may be weak or strong.
  • Acids may include, but not limited to hydrochloric or phosphoric, sulphuric, glycolic, acetic, paracetic, citric, ascorbic, and the like.
  • the solution can further comprise one or more growth factors.
  • the method can further comprise adding one or more growth factors at any stage of the method. Growth factors, for example, can comprise VEGF, bFGF, aFGF,TGFB-1 , PDGF, IGF, or any other growth factor or tissue growth stimulative agent or differentiation agent, individually or in combination, or other growth factors as described herein.
  • FIG. 8 is a flow chart illustrating an embodiment of the method 3000.
  • a scaffold is provided in a solution 3010.
  • the scaffold can be comprised of comprised of collagen, hyaluronan, or other protein, amino acid, GAG, and/or biocompatible material, and can be an allograft.
  • Growth factors can then be added to the solution 3020, followed by the addition of a base or buffer to the solution 3030 to raise the pH (make it more basic).
  • the buffer or base can comprise one or more of sodium hydroxide, water, sodium bicarbonate, phosphate buffer, sodium chloride, or calcium salt.
  • the scaffold can then be isolated after the pH is raised.
  • Growth factors for example, can comprise VEGF, bFGF, aFGF,TGFB-1 , PDGF, IGF, or any other growth factor or tissue growth stimulative agent or differentiation agent, individually or in combination, or other growth factors as described herein.
  • FIG. 9 is a flow chart illustrating an embodiment of the method 4000.
  • a medical implant is provided 4010 and energy is then provided to the medical implant 4020 to crosslink the medical implant or components thereof.
  • the medical implant can be a particulated medical implant.
  • the medical implant can be an allograft.
  • the medical implant can be an allograft that is primarily comprised of collagen.
  • the medical implant can be micronized soft tissue.
  • the energy can be heat or irradiation (gamma, ebeam, uv, and the like). In certain aspects, the energy can be irradiation of an energy of about 0.5 to about 50 kGy).
  • an additional energy can be added and the crosslinked medical implant can be terminally irradiated for the purpose of sterilization.
  • the energy added for the purpose of sterilization can be less than the initial energy added 4020.
  • the medical implant may be frozen before the step 4020. Additionally, the medical implant can be freeze-dried before or after the step 4020. Additionally, additives can be added to the medical implant before or after the step 4020. Additives can be vasodilator, a growth factor, or an amino acid, individually or in combination.
  • FIG. 10 is a flow chart illustrating an embodiment of the method 5000.
  • a mold is provided 5010, that is configured to receive a medical implant, and is configured in a desired shape (for example, which can be complementary to a soft- tissue defect of a subject).
  • a medical implant is then introduced into the mold 5015 and energy is then provided to the medical implant 5020. After the energy is provided, the medical implant can then be separated from the mold 5030.
  • the medical implant can be a particulated medical implant.
  • the medical implant can be an allograft.
  • the medical implant can be an allograft that is primarily comprised of collagen.
  • the medical implant can be micronized soft tissue.
  • the energy can be irradiation of an energy of about 2 to about 35 kGy. In certain aspects, the energy can be irradiation of an energy of about 5 to about 30 kGy. In certain aspects, the energy can be irradiation of an energy of about 10 to about 20 kGy.
  • the medical implants can comprise collagen, which can be crosslinked by the input of energy. The medical implants can comprise primarily collagen, meaning that collagen comprises the majority of the composition, or is present in an amount greater than any of the other components.
  • methods can include freezing the medical implant before providing the energy. In certain aspects, methods as described herein can include hydrating the medical implant before providing the energy. The medical implant can be placed in a mold of a desired shape before providing the energy. The medical implant can additionally be freeze dried and/or terminally irradiator (or sterilized by other sterilization means known in the art) after the application of energy.
  • one or more additives can be added to the medical implant, before providing the energy or after providing the energy.
  • the additive can be a vasodilator, for example, herparin, arginine, citrulline, or other compositions known in the art to induce nitric oxide (NO) production, a growth factor, or an amino acid.
  • Medical implants are described herein can be particulated.
  • the medical implant can be micronized soft tissue.
  • Medical implants as described herein can be made from autograft, allogeneic, or xenograft sources and may contain collagen, and growth factors/cytokines such as (but not limited to) PDGF, FGF, bFGF, aFGF, VEGF, hepatocyte growth factor (HGF), IGF, angiopoietin (ANG), ANG-2, fibronectin, TGFbl , etc.
  • growth factors/cytokines such as (but not limited to) PDGF, FGF, bFGF, aFGF, VEGF, hepatocyte growth factor (HGF), IGF, angiopoietin (ANG), ANG-2, fibronectin, TGFbl , etc.
  • Components of implants as described herein can be mixed together or layered as an injectable or structured implant.
  • Medical implants as described herein can be particulated implants, and in certain aspects can be crosslinked.
  • Medical implants described herein can be implanted surgically, injected, microneedled, and/or applied topically.
  • Medical implants described herein can be derived from follicular, dermis, fascia, amnion, amniotic fluid, placenta, umbilical cord, muscle, blood, bone marrow, or adipose tissue, their ECM, soluble proteins, or interacellular proteins.
  • medical implants as described herein can be derived from tissue that is >1 % adipose; >5% adipose; >10% adipose; >20% adipose; >30% adipose; >40% adipose; >50% adipose; >60% adipose; >70% adipose; >80% adipose; or about >90% adipose.
  • Medical implants as described herein can be particulated, gelatinized, solubilized, tissue pieces, or portions extracted.
  • the implants described herein can be combined with a delivery enhancer, flow enhancer, preservative, storage agent, protease inhibitor, stabilizer, amino acids, radioprotectant, lyoprotectant, cryoprotectant, and/or the like.
  • Medical implants as described herein can be cellular (such as cellular dermis or adipose tissue) or acellular (such as acellular dermis or adipose tissue).
  • medical implants as described herein can be refrigerated, frozen, or stored at ambient temperature. Medical implants as described herein can be dehydrated via lyophilization or supplied hydrated. Medical implants as described herein can be supplied in a syringe or a jar/bottle/vial.
  • Medical implants as described herein can be sterile filtered, tested per USP71 , or terminally sterilized via irradiation (gamma, ebeam, uv, and the like). Medical implants as described herein may be cross linked using chemical crosslinkers, heat, or irradiation (gamma, UV, ebeam, etc) to decrease degradation rate and improve volume retention.
  • Medical implants as described herein can be cleaned and disinfected using detergents, peroxides, antibiotics, water, and saline.
  • Medical implants as described herein can be cut into strips, sheets, or pieces. Medical implants can be ground or blended into fine particulate. Temperature control on cutting/grinding/blending may be used to help preserve growth factor content and prevent damage or denature proteins or other components.
  • Medical implant material (source tissue, final medical implants, or anything related to thereof or in between) may be screened/seived/filtered using syringes, needles, screens, seives, or filters.
  • Tissue implant density may be controlled by filtration, dehydration, or centrifugation speeds (100-32000 rpm/g's).
  • medical implants as described herein can comprise a backbone of one or more collagens.
  • Methods as described herein can deliver medical implants as described herein to a subject in need thereof.
  • Medical implants can be delivered to the skin of a subject in need thereof.
  • Medical implants employed in methods as described herein can be compositions comprising growth factors.
  • growth factor compositions may also contain cells (such as stem cells, keratinocytes, adipocytes, adipose derived stem cells, bone marrow derived stem cells, perivascular cells, muscle cells, stromal vascular fraction, and the like).
  • growth factor compositions as described herein can contain ascorbic acid, hemoglobin, oxygenation molecules, vasodilators, amino acids (such as arginine, lysine, methionine, cysteine, or the remaining 16 amino acids).
  • medical implants as described herein may contain adipose-derived stem cells and/or adipocytes.
  • Medical implants as described herein can be delivered to soft tissue, which in certain embodiments can be any tissue except for bone or cancellous bone.
  • medical implants can comprise bone or cancellous bone.
  • viable cells can be added to the medical implants after the medical implants are prepared.
  • methods as described herein can utilize a single injection of medical implants to an area of the in need thereof, which can be an area suffering from a soft- tissue defect, or an area where shrinkage of adipose tissue is desired.
  • the amount of medical implants which is administered to a subject can vary and can be determined by the practitioner on an individual basis according to the subject and desired outcome. Factors which can determine the amount of medical implants administered to a subject can include the degree of the soft tissue defect, and the degree to which the subject desires treatment.
  • medical implants can comprise a bioactive intracellular component.
  • a bioactive intracellular component can be a platelet-derived growth factor, a hepatocyte growth factor, an insulin growth factor, an angiopoietin, a fibronectin, a transforming growth factor, a nerve growth factor, a fibronectin, an integrin, a bone morphogenetic protein, an epidermal growth factor, an insulin-like growth factor, a fibroblast growth factor, vascular endothelial growth factor, osteoprotegerin, and osteopontin, and various combinations thereof.
  • an effective amount of a tissue implant can be an amount of tissue implant that contains a bioactive intracellular component at a concentration of at least at least 1 pg/g.
  • an effective amount of a tissue implant can be an amount of tissue implant that contains a bioactive intracellular component at a concentration of about 0 pg/g to about 100 mg/g.
  • An effective amount of a tissue implant can be an amount of tissue implant comprising a-fibroblast growth factor is present at a concentration of at least 1 pg/g.
  • An effective amount of a tissue implant can be an amount of tissue implant comprising ⁇ -fibroblast growth factor is present at a concentration of at least 1 pg/g.
  • An effective amount of a tissue implant can be an amount of tissue implant comprising vascular endothelial growth factor is present at a concentration of at least 1 pg/g.
  • An effective amount of a tissue implant can be an amount of tissue implant comprising acidic fibroblast growth factor and is present at a concentration of at least 1 pg/g.
  • An effective amount of medical implants as described herein administered to a subject in need thereof to treat soft-tissue defects can be an amount of tissue implant that contains a bioactive intracellular component at a concentration of at least at least 1 pg/mL.
  • An effective amount of medical implants as described herein administered to a subject in need thereof to treat soft-tissue defects can be an amount of tissue implant that contains a bioactive intracellular component at a concentration of at least at least 10 pg/mL.
  • An effective amount of medical implants as described herein administered to a subject in need thereof to treat soft- tissue defects can be an amount of tissue implant that contains a bioactive intracellular component at a concentration of at least at least 100 pg/mL.
  • An effective amount of medical implants as described herein administered to a subject in need thereof to treat soft-tissue defects can be an amount of tissue implant that contains a bioactive intracellular component at a concentration of at least at least 1000 pg/mL.
  • An effective amount of medical implants as described herein administered to a subject in need thereof to treat soft-tissue defects can be an amount of tissue implant that contains a bioactive intracellular component at a concentration of at least at least 10000 pg/mL.
  • An effective amount of medical implants as described herein administered to a subject in need thereof to treat soft-tissue defects can be an amount of tissue implant that contains a bioactive intracellular component at a concentration of at least at least 100000 pg/mL.
  • An effective amount of medical implants as described herein administered to a subject in need thereof to treat soft-tissue defects can be an amount of tissue implant that comprises one or more of: aFGF in an amount of at least 100,000 pg/mL; FGF in an amount of at least 100,000 pg/mL; acidic fibroblast growth factor (aFGF) in an amount of at least 100,000 pg/mL; basic fibroblast growth factor (bFGF) in an amount of at least 100,000 pg/mL; epidermal growth factor (EGF) in an amount of at least 10,000 pg/mL; hepatocyte growth factor activator (HGFa) in an amount of at least 100,000 pg/mL; hepatocyte growth factor b (HGFb) in an amount of at least 100,000 pg/mL; insulin-like growth factor 1 (IGF-1 ) in an amount of at least 10,000 pg/mL; platelet derived growth factor BB in an amount of at least 10,000 pg/mL; transforming growth factor
  • an amount effective comprises VEGF in an amount of about 5,000 pg/mL to about 1 ,000,000 pg/mL. In an embodiment, an amount effective comprises VEGF in an amount of about 66,000 pg/mL.
  • Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 0.01 cc to about 100cc. Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 0.01 cc to about 1 cc. Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 1 cc to about 10cc.
  • Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 10cc to about 100cc. Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 10cc. Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 2cc to about 9cc. Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 3cc to about 8cc. Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 4cc to about 7cc.
  • Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 5cc to about 6cc. Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 1cc to about 20cc. Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 2cc to about 19cc. Effective amounts of medical implants as described herein can be delivered to a subject in need thereof in a volume of about 5cc to about 15cc.
  • kits for treating soft-tissue defects can comprise one or more dosages of medical implants as described herein, wherein each of the one or more dosages contains an effective amount of medical implants as described herein.
  • kits can also comprise delivery devices (such as applicators or needles) according to methods and devices of the present disclosure.
  • the medical implant of the kit can be pre-loaded into the delivery device and be ready to delivered by a practitioner to a subject.
  • soft tissues which can experience defects can include bone marrow, blood, adipose, skin, muscle, vasculature, cartilage, ligament, tendon, fascia, pericardium, nerve, and hair. These soft tissues may also include organs such as the pancreas, heart, kidney, liver, intestine, and stomach. In certain aspects, as used herein soft tissue can be any tissue for example, mesodermal, endodermal, and ectodermal tissues. Examples of these tissues include bone marrow, blood, adipose, skin, muscle, vasculature, cartilage, ligament, tendon, fascia, pericardium, nerve, and hair.
  • the medical implants can be further sterilized to reduce the microorganism contamination to less than about 10 "3 microorganisms.
  • Typical sterilization methods include, but are not limited to, combinations of washing with or without pressurization, centrifugation with various chemicals such as alcohols and/or detergents, and combining antibiotics with low- dose radiation. While these processing methods reduce the amount of microorganism contamination, they also can damage the medical implant and result in the loss of many intracellular proteins and molecules.
  • a soft tissue implant contains a bioactive intracellular component of an adipose cell and a carrier substrate, where the soft tissue implant is prepared by harvesting an adipose cell from a donor, selectively lysing the adipose cell to obtain the bioactive intracellular components and combining the bioactive intracellular component with a carrier substrate.
  • the soft tissue implant can be directly administered to a subject in need thereof.
  • soft medical implants, grafts, and fillers produced by the methods described herein.
  • the methods, soft medical implants, grafts, fillers, devices, and kits described herein offer several advantages to current soft tissue grafts at least insofar as they incorporate endogenous intracellular components, while minimizing the immunogenicity of the soft tissue implant.
  • Soft tissues include, any tissue or organ that is not bone, including, but not limited to adipose tissue, muscle, cartilage, tendons, and ligaments.
  • the harvested cells are adipose cells.
  • the soft tissues can be autologous, allogeneic, xenogeneic, or syngeneic in origin. In order to minimize immunogenicity, the use of autologous cells is most advantageous. In other words, it is preferred if the harvested cells were obtained directly or indirectly (i.e. from an in vitro culture containing cells from the subject to receive the implant) from the subject that is to receive the soft tissue implant.
  • autologous adipose cells are harvested.
  • the tissue or cells are allogeneic.
  • the harvested soft tissue cells are cultured in vitro for an amount of time using suitable cell culture methods generally known in the art.
  • suitable cell culture methods generally known in the art.
  • the culture conditions will vary depending on the cell type.
  • cells from adipose tissue are cultured in vitro for about 1 day to about 6 months.
  • the cultured cells are harvested as previously described.
  • adipose cells are harvested from a donor and cultured in vitro, until harvested as previously described.
  • Medical implants as described herein can comprise growth factors, particularly vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), transforming growth factor beta 1 (TGFbl), acidic fibroblast growth factor (aFGF), insulin-like growth factor (IGF).
  • VEGF vascular endothelial growth factor
  • bFGF basic fibroblast growth factor
  • TGFbl transforming growth factor beta 1
  • aFGF acidic fibroblast growth factor
  • IGF insulin-like growth factor
  • Any given soft tissue protein and/or other bioactive factor can be present in the soluble soft tissue protein composition at a concentration of 0 ⁇ g/g to about 100 mg/g of isolated protein in the final product, dehydrated or otherwise provided.
  • soluble soft tissue protein composition can include a stabilizer composition or stabilizer compounds.
  • Suitable stabilization compounds can include, but are not limited to preservatives, antibiotics, antivirals, antifungals, pH stabilizers, osmostablizers, anti-inflammants, anti-neoplastics, chemotherapeutics, immunomodulators, chemoattractants, growth factors, anticoagulants, or combinations thereof.
  • the stabilization solution can increase shelf life of the soft tissue soluble protein composition and/or reduce denaturation of proteins during dehydration, sterilization, and/or storage.
  • other materials such as nitrogen, can be used to help reduce free radical formation and denaturation during sterilization.
  • the stabilization solution per cc of final product can be about 1 mg sucrose, about 5mg Glycine, about 3.7 mg l-Glutamic Acid, about 0.02 mg NaCI, and about 0.02 mg Polysorbate-80.
  • the stabilizer solution may contain glycerol, amino acids, polaxomers, carbomers, carbohydrates, polysaccharides, sugars, or salts.
  • the final volume of a medical implant according to compositions and methods as described herein can be at least 1 cc, or 1 cc to about 100 cc, about 1 cc to about 50 cc, 1 cc to about 25 cc, about 1 cc to about 20 cc, about 1 cc to about 10 cc.
  • the final soluble soft tissue protein product can be dehydrated or reconstituted to achieve a desired volume or particular protein concentration or composition.
  • Cells as described herein can be bone-marrow derived stem cells that are supported physiologically by a fibrous tissue called the stroma in the subject.
  • a fibrous tissue called the stroma in the subject.
  • stem cells in bone marrow There are two main types of stem cells in bone marrow: (1) hematopoietic stem cells and (2) bone marrow mesenchymal stem cells (bmMSCs).
  • bmMSCs can differentiate into a variety of cells types including without limitation, fibroblasts, chondrocytes, osteocytes, myotubes, stromal cells, adipocytes, astrocytes, and dermal cells.
  • bone marrow stroma contains other types of cells including fibroblasts (reticular connective tissue) macrophages, adipocytes, osteoblasts, osteoclasts, red blood cells, white blood cells, leukocytes, granulocytes, platelets, and endothelial cells.
  • fibroblasts reticular connective tissue
  • adipocytes adipocytes
  • osteoblasts osteoclasts
  • red blood cells white blood cells
  • leukocytes granulocytes
  • platelets granulocytes
  • endothelial cells endothelial cells
  • Proteins as described herein can be proteins derived from soluble bone marrow protein compositions that can contain proteins and/or other non-recombinant bioactive factors derived from bone marrow mesenchymal stem cells, fibroblasts, chondrocytes, osteocytes, red blood cells, white blood cells, leukocytes, granulocytes, platelets, and/or osteoclasts.
  • the proteins can be intracellular proteins or membrane associated proteins.
  • Such proteins include without limitation, bone morphogenetic proteins (BMPs) ⁇ e.g.
  • TGF- ⁇ TGF ⁇ 2
  • EGF epidermal growth factor
  • HGF hepatocyte growth factor
  • IGFs insulin-like growth factors
  • FGFs fibroblast growth factors
  • VEGF vascular endothelial growth factor
  • PDGF-BB platelet derived growth factor - BB
  • OPG osteoprotegerin
  • OPN osteopontin
  • adipose stem cells adipose stem cells, apidocytes, mesenchymal stem cells, bone marrow stromal cells, progenitor cells, etc. , that can remain viable in medical implants or in the tissue of the subject once the medical implants are implanted.
  • Example 1 Medical implants for providing cushioning or support for soft tissue defects
  • a medical implant that is at least partially comprised of collagen, hyaluronan, protein, amino acid, peptide, glycosaminoglycan (GAG), and/or biocompatible material that may cushion or support soft tissue defects such as dermis, adipose, muscle, joint, spinal disc, nerve, hair, fascia, vasculature, stroma, or other defects either created surgically, naturally occurring, or due to loss due to aging or disease (such as HIV lipoatrophy).
  • Medical implants as described herein may be derived from allograft, autograft, xenograft, or synthetic sources.
  • the medical implants as described herein can be used for joint injections (such as knee, finger, toes, ankles, hips, elbows, shoulders, jaw, or other joints), spinal disc (ie vertebral disc in a spine) injections, breast reconstruction, nipple reconstruction, foot pad atrophy, arthritis, or used subcutaneously, subdermally, intradermally, sub glandular, interglandular, inter peritoneal, or other region of the body. Medical implants as described herein may be used for any application that would benefit from support and cushioning in a subject.
  • Medical implants as described herein may be combined with cells or growth factors from other sources such as adipose, bone marrow, blood, amniotic fluid, or from another matrix such as intestine, bladder, placental, or other soft tissue.
  • sources such as adipose, bone marrow, blood, amniotic fluid, or from another matrix such as intestine, bladder, placental, or other soft tissue.
  • These other sources may be allograft, autograft, xenograft, or synthetic sources.
  • Example 2 Medical implants for the alleviation of pain
  • a medical implant as described herein (and in Example 1 above) partially comprised of allogeneic, acellular, delipidized, and decontaminated adipose, dermis, and/or fascia was injected into spinal disc and knee of patients to reduce pain and delay or eliminate the need for surgical intervention relating. After injection, patients had pain relief and did not require immediate surgery.
  • Example 3 A flowable medical implant
  • a particulated implant partially comprised of adipose, dermis, and/or fascia that has particles that are 1 mm in smallest dimension but 1.5mm or greater in a perpendicular dimension can be injected into adipose voids to correct soft-tissue defects in a subject in need thereof, defects such as rippling of breast implants, asymmetry of the breast, wrinkles in the face or neck, cellulite defects, liposuction defects, lumpectomy cavity, tumor resections, and the like.
  • Example 4 Adjusting flow of medical implants
  • a medical implant at least partially comprised of collagen, hyaluronan, protein, amino acid, peptide, glycosaminoglycan (GAG), or biocompatible material that is flowable out of a cannula.
  • the medical implant may have different flow characteristics at different temperatures or pH levels. Ph adjustment (such as pH lower than 7) can be used to decrease crosslinkinking or selfassembly of collagen, proteins, or peptides that may be present in the medical implant.
  • the implant may also be more flowable with altered pH or at temperatures above/below 37C, room temperature, or implantation temperature.
  • FIG. 11 is a graph showing energy expended on first mix.
  • Example 5 Flexible applicator for medical implant delivery
  • Described in the present example is a flexible applicator that can allow for accurate delivery of a fluid, gel, or scaffold.
  • the applicator can comprise a flexible tip, plunger/wire, crimped tube, and the like. Non-implantable portions of the applicator would be removed after placement of the implant in the desired location. Examples of possible embodiments are attached.
  • Flexible applicators as described herein can be used for the implantation of medical implants as described herein. Medical implants as described herein can be used for the treatment of soft-tissue defects in a subject, for example to fill defects in skin, adipose, muscle, vasculature, or other soft tissue void.
  • FIG. 3 is a photograph of an embodiment of a flexible applicator as described herein.
  • Described herein are medical implants that can be implanted/injected into a subject, or applied onto a subject in a location where growth factors/agents can be exposed to biocompatible scaffolding material where the combination of scaffold/growth factor solution has an acidic pH.
  • the scaffold may be acidic and/or the growth factors can be in an acidic medium.
  • the binding of those growth factors onto the scaffold can be increased by bringing the pH of the growth factors/scaffold combination to a higher pH.
  • the pH can be elevated using a buffer or a base.
  • Growth factors may consist of proteins, cytokines, peptides, amino acids, etc.
  • Growth factors/agents according to the present example may be VEGF, bFGF, aFGF, TGFB-1 , PDGF, IGF, or any other growth factor or tissue growth simulative agent or differentiation agent.
  • Scaffolds may be derived from allograft or xenograft tissue sources or created synthetically.
  • the scaffolds may be at least partially comprised of collagen, hyaluronan, or other protein, amino acid, GAG, and/or biocompatible material
  • Acids used may be weak or strong. Acids may include, but not limited to hydrochloric or phosphoric, sulphuric, glycolic, acetic, paracetic, citric, ascorbic, and the like.
  • Bases or buffers can be sodim hydroxide (NaOH), water, sodium bicarbonate, phosphate buffer, sodium chloride (NaCI), Calcium salt, or any other material that will raise pH of an acidic solution.
  • bone in an embodiment of the present example, bone can be combined with growth factors/agents with an acidic pH.
  • the pH of the bone/growth agents can then be raised by the addition of sodium bicarbonate.
  • the resulting bone is growth factor enriched and growth factors/agents are bound to the bone at a level higher than before treatment with the sodium bicarbonate.
  • adipose tissue can be combined with growth factors/agents with an acidic pH.
  • the pH of the adipose/growth agents can then be raised by the addition of sodium bicarbonate.
  • the resulting adipose is growth factor enriched and growth factors/agents are bound to the adipose at a higher level than before sodium bicarbonate.
  • Neutralization of the pH to 5 resulted in lower growth factor content than neutralization to pH 6.
  • Neutralization of the pH to 6 resulted in lower growth factor content than neutralization to pH 7.
  • Neutralization of the pH to above about 7.0 resulted in higher growth factor content than neutralization to pH less than about 7.0.
  • a scaffold containing collagen can be combined with growth factors/agents with an acidic pH.
  • the collagen/growth agents are then raised in pH by the addition of sodium bicarbonate.
  • the resulting collagen is growth factor enriched and growth factors/agents are bound to the collagen at a higher level.
  • mineralized cortical or cancellous bone In an embodiment of the present example, mineralized cortical or cancellous bone
  • the solution pH is raised by adding a buffer or base to above pH 4.
  • the growth factors and growth agents that were bound to the mineral (and loosely bound to collagen or lipid) are released into the solution and raising the pH causes those growth factors/agents to bind to the extracellular matrix of the bone.
  • mineral such as calcium
  • Example 7 Medical implant with elevated VEGF concentration
  • a medical implant derived from blood that has an elevated VEGF concentration by lysing and harvesting VEGF from the intracellular space of red blood cells.
  • the implant may be used in combination with platelets from that same donor.
  • Implant may be frozen, freeze-dried, allogeneic, autologous, or xenographic.
  • Example 8 Removing desired cell populations from medical implants
  • Implant derived from allogeneic or xenograft tissue where certain cell populations are removed and desired cell populations are retained. Implant may be cryopreserved and stored frozen until use and decontaminated using antimicrobials such as antibiotics or other non-toxic materials.
  • adipose tissue is ground very lightly to resize the particles and partially or completely kill adipocytes and treated with water to remove non-desired cell populations, for example red blood cells and white blood cells.
  • the progenitor/stem cells can still remain.
  • the implant can be treated with antibiotics such as gentamycin, vancomycin, and amphotericin B.
  • the implant can be aliquoted into individual containers and cryoprotectant is added prior to freezing.
  • Example 9 Induction of a higher volume retention of medical implants
  • a medical implant that has greater amino acid/peptide/protein/GAG content (such as collagen or hyaluronan) to induce a higher volume retention for correction of soft tissue defects/voids.
  • Particulated implants may be further concentrated by centrifugation or rehydration in smaller volumes to increase protein/GAG concentration per unit volume.
  • FIG. 12 is a graph of volume retention versus non-protein/GAG mass.
  • a medical implant that can be crosslinked and self-assembled by energy deposition (such as irradiation, light, or heat).
  • the medical implant can then be sheared to improve flowability through smaller gage needles. Shear may be introduced by rapidly cycling the flowable implant through a small aperture >5mm.
  • Example 11 Reduction of the size and appearance of adipose tissue
  • the implant can be comprised of a protein, amino acid, GAG such as chitosan.
  • the material can be injected, microneedled, or topically applied to the skin.
  • the material can contain additives to help transport into the skin, adipose, or subcutaneous layers to aid in function.
  • the material can help in the release of triglycerol, reducing the size of adipocytes, killing adipocytes, tightening the skin, or any other way to reduce the appearance of adipose tissue.
  • An enzyme can be added to the medical implant according to the present example (such as lysozyme) to help with the activity.
  • additives such as L-carnitine, ascorbic acid, caffeine, forms of glucosamine, or l-arginine may also be added to improve function.
  • FIGs. 13A and 13B are thermal images depicting before and after thermal images were taken to visualize temperature differences created by subcutaneous fat insulation.
  • FIG. 13A is before application and FIG. 13B is after application of the topical about a week later. A change on one side of the body can be seen.
  • Example 12 Crosslinking of medical implants
  • a particulated medical implant containing collagen that can be crosslinked to form a contiguous scaffold by using energy such as irradiation.
  • the medical implant may be hydrated and/or frozen during delivery of such energy.
  • the medical implant may be dehydrated and compressed for less invasive delivery.
  • the medical implant may gel or expand upon rehydration resulting in a porous shape memory implant.
  • the self-assembly or crosslinking may also effect the water content, hydrophilicity, or hydrophobicity.
  • Additives such as vasodilators, growth enhancers, or arginine may be added to help with vascularization and nutrient transport.
  • additives such as amino acids (arginine, lysine, glucosamine, and the like, natural or synthetic) may also be used to help bulk up the implant giving it a higher viscosity, G', G", compression resistance, resiliency, or the like.
  • soft tissue was micronized and poured into a mold.
  • the material was frozen and irradiated at -10, 20, 30, and 40 kGy. After irradiation, the implants were contiguous in the shape of the mold. The higher the dose of irradiation, the more stiff and stronger the shapes became.
  • Embodiments of the present disclosure will employ, unless otherwise indicated, techniques of separating, testing, and constructing materials, which are within the skill of the art. Such techniques are explained fully in the literature.

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Abstract

L'invention concerne des implants bioactifs, des procédés de fabrication et des procédés de pose. Selon certains aspects, l'invention concerne des implants bioactifs qui peuvent être utilisés pour aider à réparer des déficits en tissus mous sous-cutanés chez un sujet et/ou pour traiter la douleur chez un sujet. L'invention concerne également leurs procédés de fabrication, d'utilisation et d'administration. Les implants bioactifs peuvent être préparés en prélevant des cellules ou des tissus d'un donneur et en lysant sélectivement les cellules ou les tissus afin d'obtenir le contenu intracellulaire. L'invention concerne également des dispositifs de pose permettant de poser les implants bioactifs décrits dans la description et des kits qui comprennent les implants bioactifs décrits dans la description.
PCT/US2018/053495 2017-09-29 2018-09-28 Implants bioactifs et procédés de fabrication WO2019067942A1 (fr)

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WO2022155590A1 (fr) * 2021-01-18 2022-07-21 Advanced Biologics, Llc Implants bioactifs et méthodes de fabrication

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