Traversal of nucleic acid molecules through a fluid space and expression in repair cells

Abstract

Disclosed are methods for use in transferring nucleic acids into cells at a wound site associated with a fluid space. These gene transfer protocols are suitable for use in transferring various nucleic acids into cartilage, cardiac muscle, and other tissues, and have many uses including treating diseases such as arthritis and ischemic heart disease, and promoting wound healing. The invention further disclosed pharmaceutical compositions that may be used in the practice of the invention to transfer the nucleic acid of interest. Such compositions include any multi-partitioned biocompatible matrix in combination with multiple nucleic acids of interest.

1 . A method for transferring a nucleic acid molecule into cells associated with a fluid space, comprising contacting a wound site with a composition comprising a nucleic acid molecule and a biocompatible matrix, the wound site being situated in a tissue associated with the fluid space. 
   
   
       2 . The method of  claim 1  wherein the tissue is cartilage. 
   
   
       3 . The method of  claim 1  wherein the tissue comprises the bone/cartilage interface. 
   
   
       4 .- 8 . (canceled) 
   
   
       9 . The method of  claim 1  wherein the nucleic acid molecule is a DNA molecule. 
   
   
       10 .- 12 . (canceled) 
   
   
       13 . The method of  claim 9  wherein the DNA molecule comprising a promoter operably linked to a sequence encoding a gene product. 
   
   
       14 . The method of  claim 1  wherein the nucleic acid molecule is an RNA molecule. 
   
   
       15 .- 16 . (canceled) 
   
   
       17 . The method of  claim 1  wherein the biocompatible matrix is a biological matrix. 
   
   
       18 . (canceled) 
   
   
       19 . The method of  claim 17  wherein the biological matrix is selected from the group consisting of collagen, purified proteins, purified peptides, polysaccharides, and extracellular matrix compositions. 
   
   
       20 .- 35 . (canceled) 
   
   
       36 . The method of  claim 1  wherein the biocompatible matrix is a collagen, metal, hydroxyapatite, bioglass, aluminate, bioceramic materials, hyaluronic acid polymers, acrylic ester polymer, lactic acid polymer, glycolic acid polymer, lactic acid/glycolic acid polymer, purified proteins, purified peptides, and extracellular matrix compositions. 
   
   
       37 . The method of  claim 9  wherein the DNA molecule encodes a therapeutic protein. 
   
   
       38 . The method of  claim 37  wherein the therapeutic protein is a growth factor. 
   
   
       39 .- 43 . (canceled) 
   
   
       44 . A method of stimulating gene expression in cartilage progenitor cells located within a cartilage progenitor tissue site of an animal, comprising contacting the tissue site with a composition comprising a chondrogenic gene and a biocompatible matrix. 
   
   
       45 . The method of  claim 44  wherein expression of the gene in the cells stimulates the cells to promote cartilage tissue repair or regeneration. 
   
   
       46 .- 47 . (canceled) 
   
   
       48 . The method of  claim 44  wherein the chondrogenic gene is a parathyroid hormone (PTH) gene, a bone morphogenic protein (BMP) gene, a cartilage-derived morphogenic protein (CDMP) gene, a growth factor gene, or a growth factor receptor gene. 
   
   
       49 .- 54 . (canceled) 
   
   
       55 . The method of  claim 44  wherein the biocompatible matrix is a collagenous, polyethylene, poly(vinyl alcohol) hydrogel, polyethylene glycol, methyl cellulose, hydroxyapatite, acrylic ester polymer, lactic acid polymer, glycolic acid polymer, reconstituted fibrin-thrombin gels, anhydride polymer, orthoester polymer, hyaluronan, fibrin, carbon fiber, tetrafluoroethylene polymer, polyester, or lactic acid/glycolic acid polymer matrix. 
   
   
       56 .- 59 . (canceled) 
   
   
       60 . The method of  claim 44  wherein the cartilage progenitor tissue site is a site of cartilage injury. 
   
   
       61 .- 63 . (canceled) 
   
   
       64 . A method of stimulating cartilage repair or regeneration comprising implanting at a cartilage defective site a matrix-gene composition comprising a chondrogenic gene and a biocompatible matrix. 
   
   
       65 .- 66 . (canceled) 
   
   
       67 . A method of treating arthritis comprising implanting at a cartilage defective site a matrix-gene composition comprising a chondrogenic gene and a biocompatible matrix. 
   
   
       68 .- 72 . (canceled) 
   
   
       73 . The method of  claim 64  wherein the chondrogenic gene is a PTH gene, a BMP gene, a CDMP gene, a growth factor gene, a growth factor receptor gene. 
   
   
       74 .- 78 . (canceled) 
   
   
       79 . The method of  claim 64  wherein the biocompatible matrix is a collagenous, polyethylene, poly(vinyl alcohol) hydrogel, polyethylene glycol, methyl cellulose, hydroxyapatite, acrylic ester polymer, lactic acid polymer, glycolic acid polymer, reconstituted fibrin-thrombin gels, anhydride polymer, orthoester polymer, hyaluronan, fibrin, carbon fiber, tetrafluoroethylene polymer, polyester, or lactic acid/glycolic acid polymer matrix. 
   
   
       80 .- 87 . (canceled)