title: Targeted gene-and-host progenitor cell therapy for nonunion bone fracture repair.
abstract: Nonunion fractures present a challenge to orthopedics with no optimal solution. In-vivo DNA electroporation is a gene-delivery technique that can potentially accelerate regenerative processes. We hypothesized that in vivo electroporation of an osteogenic gene in a nonunion radius bone defect site would induce fracture repair. Nonunion fracture was created in the radii of C3H/HeN mice, into which a collagen sponge was placed. To allow for recruitment of host progenitor cells (HPCs) into the implanted sponge, the mice were housed for 10 days before electroporation. Mice were electroporated with either bone morphogenetic protein 9 (BMP-9) plasmid, Luciferase plasmid or injected with BMP-9 plasmid but not electroporated. In vivo bioluminescent imaging indicated that gene expression was localized to the defect site. Microcomputed tomography (µCT) and histological analysis of murine radii electroporated with BMP-9 demonstrated bone formation bridging the bone gap, whereas in the control groups the defect remained unbridged. Population of the implanted collagen sponge by HPCs transfected with the injected plasmid following electroporation was noted. Our data indicate that regeneration of nonunion bone defect can be attained by performing in vivo electroporation with an osteogenic gene combined with recruitment of HPCs. This gene therapy approach may pave the way for regeneration of other skeletal tissues.
authors.affiliation: Skeletal Biotech Laboratory, Hebrew University-Hadassah Medical Center, Faculty of Dental Medicine, Jerusalem, Israel.
authors.forename: Nadav
authors.initials: N
authors.lastname: Kimelman-Bleich
authors.forename: Gadi
authors.initials: G
authors.lastname: Pelled
authors.forename: Yoram
authors.initials: Y
authors.lastname: Zilberman
authors.forename: Ilan
authors.initials: I
authors.lastname: Kallai
authors.forename: Olga
authors.initials: O
authors.lastname: Mizrahi
authors.forename: Wafa
authors.initials: W
authors.lastname: Tawackoli
authors.forename: Zulma
authors.initials: Z
authors.lastname: Gazit
authors.forename: Dan
authors.initials: D
authors.lastname: Gazit
mesh_terms: D000818:Animals; D001861:Bone Regeneration / Q000235:genetics*; D003094:Collagen / Q000008:administration & dosage; D018274:Electroporation / Q000379:methods*; D005260:Female; D005599:Fractures, Ununited / Q000473:pathology / Q000628:therapy*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D055427:Growth Differentiation Factor 2 / Q000096:biosynthesis / Q000235:genetics*; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008809:Mice, Inbred C3H; D010012:Osteogenesis / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D013234:Stem Cells / Q000502:physiology*; D014945:Wound Healing / Q000235:genetics
chemical_list: D055427:Growth Differentiation Factor 2; D003094:Collagen; D008156:Luciferases