Compositions and methods for augmentation and regeneration of living tissue in a subject

Abstract

The present invention provides for a composition, for augmentation and regeneration of living tissue in a subject, comprising a population of porous microparticles of a biodegradable polymer, one or more mammalian cell populations, and optionally, a biocompatible adhesive.

1 . A composition comprising a population of porous microparticles of a biodegradable polymer. 
     
     
         2 . A composition according to  claim 1 , wherein the porosity is 50 to 95%. 
     
     
         3 . A composition according to  claim 1 , wherein the size of the microparticles is between 20-110 μm. 
     
     
         4 . A composition according to  claim 1  for augmentation and regeneration of living tissue in a subject. 
     
     
         5 . A composition according to  claim 1  further comprising one or more mammalian cell populations. 
     
     
         6 . A composition according to  claim 1  further comprising a biocompatible adhesive. 
     
     
         7 . The composition according to  claim 1 , wherein one or more populations of a mammalian cell is attached to said population of microparticles of a biodegradable polymer. 
     
     
         8 . A composition according to  claim 1 , wherein the microparticles are in the form of microspheres of the biodegradable polymer. 
     
     
         9 . A composition according to  claim 1 , wherein the microparticles are in the form of irregular shapes, such as flakes of the biodegradable polymer. 
     
     
         10 . A composition according to  claim 1 , wherein said biodegradable polymer comprises or consists of a polymer selected from the group consisting of: a) Homo- or copolymers of : glycolide, such as L-lactide, DL-lactide, meso-lactide (polylactide, PLA), e-caprolactone (polycapro lactone, PCL), 1,4-dioxane-2-one, d-valerolactone, B-butyrolactone, g-butyrolactone, e-decalactone, 1,4-dioxepane-2-one, 1,5-dioxepan-2-one, 1,5,8,12-tetraoxacyclotetradecane-7-14-dione, 1,5-dioxepane-2-one, 6,6-dimethyl-1,4-dioxane-2-one, and trimethylene carbonate; b) Block-copolymers of mono- or difunctional polyethylene glycol and polymers of a) mentioned above; c) Block copolymers of mono- or difunctional polyalkylene glycol and polymers of a) mentioned above; d) Blends of the above mentioned polymers; and e) polyanhydrides and polyorthoesters; such as copolymers of poly(D,L-lactide-co-glycolide) (PLGA), MPEG-PLGA (methoxypolyethyleneglycol)-poly(D,L-lactide-co-glycolide). 
     
     
         11 . A composition according to  claim 1  wherein the biodegradable polymer consists or comprises PLGA or MPEG-PLGA. 
     
     
         12 . A composition according to  claim 1 , wherein said microparticles are prepared by freeze drying a solution comprising the biodegradable polymer in solution. 
     
     
         13 . A composition according to  claim 1 , wherein said microparticles are prepared by ultrasonic atomisation. 
     
     
         14 . A composition according to  claim 5 , wherein said cells are autologous, homologus (allogenic) or xenogenic in origin relative to cells of said living tissue in a subject. 
     
     
         15 . The composition according to  claim 1 , wherein the concentration of cells in the composition is about 0.1×10 4  cells to about 10×10 6  cells per ml. 
     
     
         16 . The composition according to  claim 1  further comprising a component which facilitates the cell adhesion and/or in-growth for regeneration of tissue to biodegradable polymer microparticles, such as a component selected from the group consisting of: chondroitin sulfate, hyaluronan, hyaluronic acid (HA), heparin sulfate, heparan sulfate, dermatan sulfate, growth factors, fibrin, fibronectin, elastin, collagen, such as collagen type I and/or type II, gelatin, and aggrecan, or any other suitable extracellular matrix component. 
     
     
         17 . The composition according to  claim 1 , further comprising a compound in the synthetic biodegradable scaffold, wherein said further compound is a growth factors, such as Insulin-like growth factor 1 (IGF-1), or transforming growth factors (TGFs), such as TGF-alpha or TGF-beta, or FGFs, such as FGF-1 or FGF-2 or bone morphogenic protein (BMP). 
     
     
         18 . The composition according to  claim 1  wherein hyaluronic acid and/or dermatan sulphate is incorporated into the biodegradable polymer microparticles. 
     
     
         19 . The composition according to  claim 1 , wherein at least some of the microparticles comprises hydroxyapatite and/or calcium phosphate. 
     
     
         20 . A method for the preparation of a population of microparticles, preferably with a regular microparticle structure, such as microspheres, said method comprising:
 a) Preparing a solution of the polymer in a solvent;   b) Admixing the solution formed in step a) with a non-solvent;   c) Atomise the solution formed in step b) into a non-solvent at below room temperature (preferably below 0° C.) to form the microparticles;   d) Collect the particles formed, preferably by filtration;   e) Optionally dry particles, preferably by application of a vacuum or by freeze drying.   
     
     
         21 . A method for the preparation of a population of microparticles, preferably with an irregular microparticle structure, said method comprising:
 a) Preparing a solution of the polymer in a solvent;   b) Admixing the solution formed in step a) with a non-solvent;   c) Freeze dry the admixed solution formed in step b);   d) Collect the particles formed;   e) Optionally size fractionate particles, preferably by sieving.   
     
     
         22 . A method according to  claim 21 , wherein the solvent in a) is selected from the group consisting of 1,4-dioxane, dimethylcarbonate, or 1,3-dioxolane. 
     
     
         23 . A method for the preparation of a population of microparticles, preferably with an irregular structure, said method comprising:
 a) Preparing a solution of the polymer in a solvent;   b) Atomise, for example by ultrasonic atomisation, the solution formed in step b) into a non-solvent at below room temperature (preferably below 0° C.) to form the microparticles;   c) Collect the particles formed, preferably via filtration;   d) Optionally dry particles, preferably by application of a vacuum or by freeze drying.   
     
     
         24 . A method for the preparation of a population of microparticles, said method comprising:
 a. Preparing a solution of the polymer in a solvent that can be freeze dried;   b. Atomise, for example by ultrasonic atomisation, the solution formed in step a) into a cold chamber, thereby freezing the drops to particles;   c. collect the particles formed,   d. dry particles by freeze drying.   
     
     
         25 . The method according to  claim 20 , wherein the polymer is the biodegradable polymer. 
     
     
         26 . The method according to  claim 23 , wherein the solvent is selected from the group consisting of chloroform, 1,4-dioxane, acetone, methylacetate etc, butanone, dichloromethane, and 1,3-dioxolane. 
     
     
         27 . The method according to  claim 20 , wherein the non-solvent selected from the group consisting of: lower alcohols, diethyl ether, diisopropyl ether, hexane, alkanes, and cycloalkanes. 
     
     
         28 . The microparticle population prepared by the method of  claim 20 . 
     
     
         29 . A population of microparticles of MPEG-PLGA. 
     
     
         30 . A population according to  claim 29 , wherein the particles a porous with a porosity of 50 to 95%. 
     
     
         31 . A population according to  claim 29 , wherein the size of the microparticles is between 20-110 μm. 
     
     
         32 . A kit comprising
 a) Porous, microparticles of a biodegradable polymer in a concentration to form a liquid;   b) A device for placing the cells.   
     
     
         33 . A kit of  claim 32 , wherein the device for placing the cells is a syringe with a needle. 
     
     
         34 . The kit of  claim 32 , further comprising cells in conc. of 1 mill cells per 50-100 mg. 
     
     
         35 . The kit of  claim 32  for repair of a muscle. 
     
     
         36 . The kit of  claim 35 , for repair of a spincter muscle. 
     
     
         37 . A kit comprising
 a) Porous, microparticles of a biodegradable polymer in a concentration to form a paste;   b) A device for placing the cells;   c) tissue glue.   
     
     
         38 . A kit of  claim 37 , wherein the device for placing the cells is a syringe without a needle, but with a flexible outlet. 
     
     
         39 . The kit of  claim 37 , further comprising cells in conc. of 1 mill pr cm 2 . 
     
     
         40 . The kit of  claim 37  for cartilage repair. 
     
     
         41 . A method for the regeneration or augmentation of a living tissue within a patient, said method comprising preparing the composition according to  claim 1  and administering said composition into the living tissue to be regenerated or augmented, such as the living tissue referred to in any one of the preceding claims. 
     
     
         42 . The method according to  claim 41 , wherein the composition is administered to the tissue via an injection. 
     
     
         43 . The method according to  claim 41 , wherein the living tissue is a muscle tissue, and wherein the one or more population of a mammalian cell comprises a population of myoblast cells, or stem cells capable of differentiation into myoblast cells. 
     
     
         44 . The method according to  claim 43 , wherein said method is a method for the treatment of a disease related to uro-gynaecological disorders, such as urinary incontinence, and the living tissue is a sphincter muscle, such as the urinary sphincter muscle. 
     
     
         45 . The method according to  claim 41 , wherein the living tissue is a cartilage tissue, and wherein the one or more population of a mammalian cell comprises a population of cells selected from the list consisting of chondrocytes, such as human articular chodrocytes, stem cells or equivalent cells capable of transformation into a chondrocyte, such as mesenchymal stem cells. 
     
     
         46 . The method according to  claim 41 , wherein the living tissue is a bone tissue, and wherein the one or more population of a mammalian cell comprises a population of osteoblasts or stem cells which are capable of differentiating into osteoblasts, such as mesenchymal stem cells. 
     
     
         47 . A composition according to  claim 1  for use as a medicament. 
     
     
         48 . A composition according to  claim 1  for use in the treatment of a disease related to uro-gynaecological disorders such as urinary incontinence, pelvic organ prolapse, and anal incontinence. 
     
     
         49 . A composition according to  claim 1  for use in the treatment of a disease related to a cartilage defect. 
     
     
         50 . A composition according to  claim 1  for use in the treatment of a disease related to a bone defect or disease.