Acellular soft tissue-derived matrices and methods for preparing same

Abstract

An acellular soft tissue-derived matrix includes a collagenous tissue that has been delipidated and decellularized. Adipose tissue is among the soft tissues suitable for manufacturing an acellular soft tissue-derived matrix. Exogenous tissuegenic cells and other biologically-active factors may be added to the acellular matrix. The acellular matrix may be provided as particles, a slurry, a paste, a gel, or in some other form. The acellular matrix may be provided as a three-dimensional scaffold that has been reconstituted from particles of the three-dimensional tissue. The three-dimensional scaffold may have the shape of an anatomical feature and serve as a template for tissue repair or replacement. A method of making an acellular soft tissue-derived matrix includes steps of removing lipid from the soft tissue by solvent extraction and chemical decellularization of the soft tissue.

We claim: 
     
         1 . A composition, comprising a delipidated, decellularized adipose tissue. 
     
     
         2 . The composition of  claim 1 , wherein said adipose tissue is substantially free of native lipids. 
     
     
         3 . The composition of  claim 1 , wherein said adipose tissue is substantially free of native cellular components. 
     
     
         4 . The composition of  claim 1 , wherein said adipose tissue is substantially free of native nucleic acids. 
     
     
         5 . The composition of  claim 1 , further comprising exogenous tissuegenic cells. 
     
     
         6 . The composition of  claim 5 , wherein said exogenous tissuegenic cells include exogenous stem cells. 
     
     
         7 . The composition of  claim 5 , wherein said exogenous tissuegenic cells include adipose-derived stem cells. 
     
     
         8 . The composition of  claim 1 , further comprising at least one exogenous growth-inductive substance. 
     
     
         9 . The composition of  claim 8 , wherein said at least one exogenous growth-inductive substance is selected from the group consisting of bone morphogenic proteins, vascular endothelial growth factor, basic fibroblast growth factor, transforming growth factor beta, platelet-derived growth factor, and neural epidermal growth-factor-like 1. 
     
     
         10 . The composition of  claim 1 , wherein said delipidated, decellularized adipose tissue is substantially free of substances that pose a significant risk of causing an immunogenic response in a patient receiving said composition. 
     
     
         11 . The composition of  claim 1 , further comprising a carrier for said delipidated, decellularized adipose tissue. 
     
     
         12 . The composition of  claim 11 , wherein said carrier includes a substance selected from the group consisting of an isotonic solution, a sodium chloride solution, lactated Ringer's solution, a phosphate-buffered saline solution, platelet rich plasma, hyaluronic acid, a derivative of hyaluronic acid, sodium hyaluronate, thrombin, fibrin, glycerin, collagen, lecithin, sugars, polysaccharides, and solutions thereof. 
     
     
         13 . The composition of  claim 11 , wherein said carrier includes a substance selected from the group consisting of polysaccharides, nucleic acids, carbohydrates, proteins, polypeptides, poly(α-hydroxy acids), poly(lactones), poly(amino acids), poly(anhydrides), poly(orthoesters), poly(anhydride-co-imides), poly(orthocarbonates), poly(α-hydroxy alkanoates), poly(dioxanones), poly(phosphoesters), poly(L-lactide) (PLLA), poly(D,L-lactide) (PDLLA), polyglycolide (PGA), poly(lactide-co-glycolide (PLGA), poly(L-lactide-co-D, L-lactide), poly(D,L-lactide-co-trimethylene carbonate), polyhydroxybutyrate (PHB), poly(ε-caprolactone), poly(δ-valerolactone), poly(γ-butyrolactone), poly(caprolactone), polyacrylic acid, polycarboxylic acid, poly(allylamine hydrochloride), poly(diallyldimethylammonium chloride), poly(ethyleneimine), polypropylene fumarate, polyvinyl alcohol, polyvinylpyrrolidone, polyethylene, polymethylmethacrylate, carbon fibers, poly(ethylene glycol), poly(ethylene oxide), poly(vinyl alcohol), poly(vinylpyrrolidone), poly(ethyloxazoline), poly(ethylene oxide)-co-poly(propylene oxide) block copolymers, poly(ethylene terephthalate)polyamide, and copolymers thereof 
     
     
         14 . A delipidated, decellularized adipose tissue. 
     
     
         15 . A method of preparing an adipose tissue-derived matrix, comprising the step of decellularizing a delipidated adipose tissue. 
     
     
         16 . A method of preparing an adipose tissue-derived matrix, comprising the separate steps of delipidating the adipose tissue and decellularizing the delipidated adipose tissue. 
     
     
         17 . A method of preparing an adipose tissue-derived matrix, comprising the steps of:
 obtaining an adipose tissue;   reducing the size of the adipose tissue;   delipidating the mechanically reduced adipose tissue with a polar organic solvent;   decellularizing the delipidated adipose tissue with a detergent solution; and   disinfecting the delipidated, decellularized adipose tissue.   
     
     
         18 . The method of  claim 17 , including the further steps of drying the disinfected adipose tissue; and reducing the dried adipose tissue to a dried particulate form. 
     
     
         19 . The method of  claim 17 , wherein said step of reducing the size of the adipose tissue includes a step of grinding the adipose tissue. 
     
     
         20 . The method of  claim 19 , wherein said step of grinding the adipose tissue includes a step of grinding the adipose tissue with a coarse grinding plate, followed by a step of grinding the ground adipose tissue with a fine grinding plate. 
     
     
         21 . The method of  claim 19 , including the further step of mechanically separating a lipid layer from the ground adipose tissue after said grinding step. 
     
     
         22 . The method of  claim 17 , wherein said polar organic solvent is selected from a group of polar solvents consisting of a polar solvent having one carbon atom, a polar solvent having two carbon atoms, a polar solvent having three carbon atoms, a polar solvent having four carbon atoms, a polar solvent having five carbon atoms, and a polar solvent having six carbon atoms. 
     
     
         23 . The method of  claim 17 , wherein said polar organic solvent includes an alcohol. 
     
     
         24 . The method of  claim 17 , wherein said polar organic solvent is selected from the group consisting of a paraffin, an aromatic hydrocarbon, a cyclic hydrocarbon, a chlorinated hydrocarbon, a fluorinated hydrocarbon, a chlorinated methane, a fluorinated methane, an alcohol, an ether, a ketone, an aldehyde, an ester, an organic acid, and combinations thereof. 
     
     
         25 . The method of  claim 17 , wherein said delipidating step includes the step of washing the mechanically reduced adipose tissue with the polar organic solvent. 
     
     
         26 . The method of  claim 17 , wherein said delipidating step includes the step of blending the mechanically reduced adipose tissue with the polar organic solvent. 
     
     
         27 . The method of  claim 17 , wherein said delipidating step includes the step of homogenizing the mechanically reduced adipose tissue with the polar organic solvent. 
     
     
         28 . The method of  claim 17 , wherein said decellularizing step includes a step of washing the delipidated adipose tissue with a detergent solution. 
     
     
         29 . The method of  claim 25 , wherein the detergent solution includes at least one of deoxycholic acid or a deoxycholate salt. 
     
     
         30 . The method of  claim 25 , wherein the detergent solution has a detergent concentration in the range of 0.1% to 5.0%. 
     
     
         31 . The method of  claim 25 , wherein the detergent solution includes a mixture of water and alcohol. 
     
     
         32 . The method of  claim 31  wherein the alcohol is present in the detergent solution in an amount in the range of about 20% to about 40% by volume. 
     
     
         33 . The method of  claim 31  wherein the alcohol is present in the detergent solution in an amount in the range of about 60% to about 80% by volume. 
     
     
         34 . The method of  claim 17 , further including the step of contacting the delipidated, decellularized adipose tissue with a solution having an acidic pH so as to form a gel. 
     
     
         35 . The method of  claim 34 , wherein said contacting step includes the step of blending the delipidated, decellularized adipose tissue with the acidic solution. 
     
     
         36 . The method of  claim 17 , further including the step of blending the disinfected adipose tissue in a polar liquid, whereby the disinfected adipose liquid and the polar liquid form a flowable gel. 
     
     
         37 . The method of  claim 17 , wherein said disinfecting step includes the step of contacting the delipidated, decellularized adipose tissue with a disinfectant solution. 
     
     
         38 . The method of  claim 32 , wherein said contacting step includes the step of soaking the delipidated, decellularized adipose tissue in the disinfectant solution. 
     
     
         39 . The method of  claim 17 , wherein said disinfecting step includes the step of blending the delipidated, decellularized adipose tissue with the disinfectant solution. 
     
     
         40 . The method of  claim 39 , wherein the disinfecting solution is an acidic solution. 
     
     
         41 . The method of  claim 40 , wherein the disinfecting solution includes peracetic acid. 
     
     
         42 . The method of  claim 37 , wherein the disinfecting solution includes an alcohol. 
     
     
         43 . The method of  claim 42 , wherein the disinfecting solution further includes a glycol. 
     
     
         44 . The method of  claim 37 , wherein said disinfecting step includes the step of washing the disinfected adipose tissue with a buffer having a physiologic pH after said contacting step. 
     
     
         45 . The method of  claim 17 , wherein said drying step includes the step of drying the disinfected adipose tissue in air. 
     
     
         46 . The method of  claim 17 , wherein said drying step includes the step of drying the disinfected adipose tissue with heat. 
     
     
         47 . The method of  claim 17 , wherein said drying step includes the step of lyophilizing the disinfected adipose tissue. 
     
     
         48 . The method of  claim 17 , wherein the step of reducing the size of the dried adipose tissue includes the step of milling the dried adipose tissue, thereby producing a dried particulate material. 
     
     
         49 . The method of  claim 17 , including the further step of packaging the dried particulate material in a dried particulate form. 
     
     
         50 . The method of  claim 17 , wherein the step of reducing the size of the dried adipose tissue includes the step of freezer-milling the dried adipose tissue, thereby producing a frozen dried particulate material. 
     
     
         51 . The method of  claim 50 , including the further step of thawing the frozen dried particulate material. 
     
     
         52 . The method of  claim 51 , including the further step of packaging the thawed dried particulate material in a dried particulate form. 
     
     
         53 . The method of  claim 18 , including the further steps of rehydrating the dried particulate material and mixing the rehydrated particulate material with a carrier. 
     
     
         54 . The method of  claim 18 , including the further steps of rehydrating the dried particulate material and seeding the rehydrated particulate material with exogenous tissuegenic cells to form a seeded scaffold material. 
     
     
         55 . The method of  claim 54 , including the further step of packaging the seeded scaffold material in a cryopreservative. 
     
     
         56 . A method of preparing an adipose tissue-derived matrix, comprising the step of processing an adipose tissue to remove substantially all lipid and cellular material from the adipose tissue, said processing step including the step of contacting the adipose tissue with a polar organic solvent, and the step of contacting the adipose tissue with a detergent solution after said step of contacting the adipose tissue with a polar organic solvent.