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 .- 87 . (canceled) 
     
     
         88 . 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, wherein repair cells associated with the fluid space are exposed to the composition comprising the nucleic acid molecule and a biocompatible matrix, which method does not involve creating or locating a perforation in a subchondral bone of a joint. 
     
     
         89 . The method of  claim 88  wherein the tissue is cartilage. 
     
     
         90 . The method of  claim 88  wherein the tissue comprises the bone/cartilage interface. 
     
     
         91 . The method of  claim 88  wherein the wound is induced by injury. 
     
     
         92 . The method of  claim 88  wherein the wound is induced by a disease state. 
     
     
         93 . The method of  claim 88  wherein the wound is an iatrogenic wound. 
     
     
         94 . The method of  claim 88  wherein the nucleic acid molecule is a DNA molecule. 
     
     
         95 . The method of  claim 94  wherein the DNA molecule comprises a promoter operably linked to a nucleic acid sequence encoding a gene product. 
     
     
         96 . The method of  claim 88  wherein the nucleic acid molecule is an RNA molecule. 
     
     
         97 . The method of  claim 88  wherein the biocompatible matrix is a biological matrix. 
     
     
         98 . The method of  claim 97  wherein the biological matrix comprises a polymer. 
     
     
         99 . The method of  claim 98  wherein the polymer is selected from a group consisting of polyethylene glycols and their derivatives, polyesters, polyethers, polyanhydrides, polyalkylcyanoacrylates, polyacrylamides, polyortheosters, polyphospazenes, polyvinylacectates, block copolymers, polytetrafluoroethylene (PTFE), and polyurethanes. 
     
     
         100 . The method of  claim 97  wherein the biological matrix is selected from the group consisting of 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, polysaccharides, and extracellular matrix compositions. 
     
     
         101 . The method of  claim 94  wherein the DNA molecule encodes a therapeutic protein. 
     
     
         102 . The method of  claim 101  wherein the therapeutic protein is a growth factor. 
     
     
         103 . The method of  claim 102  wherein the growth factor is a transforming growth factor (TGF), a fibroblast growth factor (FGF), a platelet derived growth factor (PDGF), an insulin like growth factor (IGF), hepatocyte growth factor (HGF), an epidermal growth factor (EGF), a connective tissue growth factor (CTGF), a bone morphogenic factor (BMP), keratinocyte growth factor (KGF), vascular endothelial growth factor (VEGF), or cartilage-derived morphogenic protein (CDMP). 
     
     
         104 . The method of  claim 101  wherein the therapeutic protein is selected from the group consisting of latent TGF-β binding protein (LTBP), Factor VIII, Factor IX, erythropoietin (EPO), tissue plaminogen activator (TPA), leukemia inhibitory factor (LIF), parathyroid hormone-related peptide (PTHrP), activin, inhibin, interleukin, macrophage-colony stimulating factor (M-C SF), granulocyte macrophage-colony stimulating factor (GM-CSF), skeletal growth factor (SGF), chondromodulin, mono or polyclonal antibodies and fragments thereof, enzymes involved in the production and/or processing of collagen, enzymes involved in the production and/or processing of hyalronic acid, transcription factors that trigger proliferation, differentiation, and morphogenic pathways, cell survival factors, or cell death factors. 
     
     
         105 . The method of  claim 101  wherein the therapeutic protein is a hormone. 
     
     
         106 . The method of  claim 105  wherein the hormone is a growth hormone. 
     
     
         107 . The method of  claim 105  wherein the hormone is a human parathyroid hormone (PTH).