Method for in situ repair of injured, damaged, diseased or aged articular cartilage

Abstract

A method for treatment of injured, damaged, diseased or aged articular cartilage using neo-cartilage constructs implanted into a joint cartilage lesion in situ. The implantation of the construct initiates and achieves incorporation of neo-cartilage into a native surrounding cartilage including a formation of a new superficial cartilage layer overgrowing and sealing the lesion in the joint cartilage.

1 - 20 . (canceled)  
   
   
       21 . A method for treatment and regeneration of injured, damaged, diseased or aged articular cartilage comprising steps: 
 a) obtaining about 50-4000 mg of articular hyaline cartilage;    b) isolating chondrocytes;    c) expanding said chondrocytes in culture;    d) seeding said chondrocytes into a support matrix;    e) generating a neo-cartilage construct by subjecting said seeded support matrix construct to an algorithm comprising cyclic or constant hydrostatic pressure from about 0.1 to about 10 MPa at frequency of about 0.1 to about 1 Hz for about 1 to about 8 hours per day, followed by about 16 to 23 hours of a recovery period at constant atmospheric pressure for from about 4 to about 100 days period of time at a perfusion flow rate of a culture medium of about 1 to about 500 μL in the presence of about 1% to about 20% oxygen; and    f) implanting said construct of step e) into an articular cartilage lesion.    
   
   
       22 . The method fo  claim 21  additionally comprising a step of covering said construct implanted into said lesion with a biologically acceptable top sealant deposited over the implanted neo-cartilage construct said sealant overlaying the implanted neo-cartilage construct and adjacent hyaline cartilage.  
   
   
       23 . The method of  claim 22  additionally comprising a step of depositing a biologically acceptable bottom sealant at a bottom of the lesion.  
   
   
       24 . The method fo  claim 23  wherein said top or bottom sealant is selected from the group consisting of gelatin, a copolymer of polyethylene glycol and poly-lactide or poly-glycolide, periodate-oxidized gelatin, 4-armed pentaerythritol thiol and a polyethylene glycol diacrylate, 4-armed tetra-succinimidyl ester or tetra-thiol derivatized PEG, photo-polymerizable polyethylene glycol-co-poly(α-hydroxy acid) diacrylate macromer, 4-armed polyethylene glycols derivatized with succinimidyl ester and thiol plus methylated collagen, hydrogel, derivatized polyethylene glycol (PEG), derivatized polyethylene glycol (PEG) cross-linked with alkylated collagen, tetra-hydrosuccinimidyl or tetra-thiol derivatized PEG, cross-linked PEG with methylated collagen and a combination thereof.  
   
   
       24 . The method of  claim 23  wherein the sealant is cross-linked PEG with methylated collagen.  
   
   
       25 . The method of  claim 23  wherein said top and bottom sealants are the same or different.  
   
   
       26 . The method of  claim 21  wherein the said chondrocytes obtained in step a) are non-osteoarthritic or osteoarthritic chondrocytes obtained from a patient during arthroscopy; 
 wherein the chondrocytes are isolated by a protease or a lyase, either sequentially or in a mixture thereof; and    wherein the chondrocytes are expanded by incubation in a culture medium, suspended in a gel or sol-gel solution or in a thermo-reversible gelation hydrogel (TRGH) and seeded into the support matrix of step (d).    
   
   
       27 . The method of  claim 26  wherein said support matrix is a sponge, honeycomb lattice, porous scaffold or hydrogel.  
   
   
       28 . The method of  claim 27  wherein said support matrix is prepared from a material selected from the group consisting of a Type I collagen, a Type II collagen, a Type IV collagen, a cell-contracted collagen containing a proteoglycan, a cell-contracted collagen containing a glycosaminoglycan, a cell-contracted collagen-containing a glycoprotein, gelatin, agarose, hyaluronin, fibronectin, laminin, a bioactive peptide growth factor, a cytokine, elastin, fibrin, a synthetic polymeric fiber made of a polylactic acid, a synthetic polymeric fiber made of polyglycolic, a synthetic polymeric fiber made of polyamino acid, polycaprolactone, a polyamino acid, a polypeptide gel, a polymeric thermo-reversible gelation hydrogel (TRGH), a copolymer thereof and a combination thereof.  
   
   
       29 . The method of  claim 28  wherein said chondrocytes are seeded into said matrix in the gel, sol-gel suspension or TRGH.  
   
   
       30 . The method of  claim 29  wherein said matrix is TRGH and said chondrocytes are suspended into said TRGH at a temperature below about 30° C. when the TRGH is in a liquid sol form, and wherein said TRGH is subsequently converted into a solid gel by increasing the temperature from above about 30° to about 37° C.  
   
   
       31 . The method of  claim 30  wherein said seeded matrix is subjected to the algorithm of step (e).  
   
   
       32 . The method of  claim 31  wherein said algorithm comprises cyclic hydrostatic pressure of about 3.0 MPa at frequency of about 0.1 Hz or a constant hydrostatic pressure from zero to about 3 MPa applied for about 7 days preceded or followed by the static atmospheric pressure applied for about 12 days wherein said perfusion flow rate is about 5 μL and wherein said perfusion is performed in the presence of about 2% to about 5% oxygen.  
   
   
       33 . The method of  claim 32  wherein the neo-cartilage construct comprises a suspension of chondrocytes in TRGH.  
   
   
       34 . The method of  claim 32  wherein said neo-cartilage construct is implanted into said cartilage into a cavity formed between said top sealant layer and said bottom sealant layer.  
   
   
       35 . The method of  claim 32  wherein said support matrix is a sponge or honeycomb lattice incorporated with chondrocytes and said neo-cartilage construct is implanted above the bottom sealant into said lesion after the deposition of the bottom sealant and before the deposition of the top sealant.