BMP2/7 heterodimer expression by adenovirus can stimulate bone formation at subcutaneous sites. In the present study, we evaluate whether this approach will also promote healing of cranial defects. Adenovirus expressing BMP2 or BMP7 (AdBMP2, AdBMP7) was titrated to yield equivalent BMP protein levels after transduction into murine BLK cells. Analysis of conditioned medium showed that BMP2/7 heterodimers have enhanced ability to stimulate alkaline phosphatase and Smad 1,5,8 phosphorylation relative to equivalent amounts of BMP2 or BMP7 homodimers. To measure bone regeneration, we implanted virally transduced BLK cells into critical-sized calvarial defects generated in C57BL6 mice. AdBMP2/7-transduced cells were more effective in healing cranial defects than were cells individually transduced with AdBMP2 or BMP7. Dramatic increases in bone volume fraction, as measured by microCT, as well as fusion of regenerated bone with the defect margins were noted. Thus, the use of gene therapy to express heterodimeric BMPs is a promising potential therapy for healing craniofacial bones.
Get full access to this article
View all access options for this article.
References
1.
Aoki H, Fujii M, Imamura T, Yagi K, Takehara K, Kato M, et al. -2001-. Synergistic effects of different bone morphogenetic protein type I receptors on alkaline phosphatase induction. J Cell Sci114-Pt 8-:1483–1489.
2.
Aono A, Hazama M, Notoya K, Taketomi S, Yamasaki H, Tsukuda R, et al. -1995-. Potent ectopic bone-inducing activity of bone morphogenetic protein-4/7 heterodimer. Biochem Biophys Res Commun210:670–677.
3.
Cochran DL, Jones AA, Lilly LC, Fiorellini JP, Howell H -2000-. Evaluation of recombinant human bone morphogenetic protein-2 in oral applications including the use of endosseous implants: 3-year results of a pilot study in humans. J Periodontol71:1241–1257.
4.
Franceschi RT -2005-. Biological approaches to bone regeneration by gene therapy. J Dent Res84:1093–1103.
5.
Franceschi RT, Yang S, Rutherford RB, Krebsbach PH, Zhao M, Wang D -2004-. Gene therapy approaches for bone regeneration. Cells Tissues Organs176:95–108.
6.
Geesink RG, Hoefnagels NH, Bulstra SK -1999-. Osteogenic activity of OP-1 bone morphogenetic protein -BMP-7- in a human fibular defect. J Bone Joint Surg Br81:710–718.
7.
Giannobile WV, Ryan S, Shih MS, Su DL, Kaplan PL, Chan TC -1998-. Recombinant human osteogenic protein-1 -OP-1- stimulates periodontal wound healing in class III furcation defects. J Periodontol69:129–137.
8.
Govender S, Csimma C, Genant HK, Valentin-Opran A, Amit Y, Arbel R, et al. -2002-. Recombinant human bone morphogenetic protein-2 for treatment of open tibial fractures: a prospective, controlled, randomized study of four hundred and fifty patients. J Bone Joint Surg Am84-A-:2123–2134.
9.
Hassel S, Schmitt S, Hartung A, Roth M, Nohe A, Petersen N, et al. -2003-. Initiation of Smad-dependent and Smad-independent signaling via distinct BMP-receptor complexes. J Bone Joint Surg Am85-A--Suppl 3-:44–51.
10.
Hogan BL -1996-. Bone morphogenetic proteins: multifunctional regulators of vertebrate development. Genes Dev10:1580–1594.
11.
Huang YC, Kaigler D, Rice KG, Krebsbach PH, Mooney DJ -2005-. Combined angiogenic and osteogenic factor delivery enhances bone marrow stromal cell-driven bone regeneration. J Bone Miner Res20:848–857.
12.
King GN, King N, Cruchley AT, Wozney JM, Hughes FJ -1997-. Recombinant human bone morphogenetic protein-2 promotes wound healing in rat periodontal fenestration defects. J Dent Res76:1460–1470.
13.
Kirker-Head CA -2000-. Potential applications and delivery strategies for bone morphogenetic proteins. Adv Drug Deliv Rev43:65–92.
14.
Koh JT, Ge C, Zhao M, Wang Z, Krebsbach PH, Zhao Z, et al. -2006-. Use of a stringent dimerizer-regulated gene expression system for controlled BMP2 delivery. Mol Ther14:684–691.
15.
Krebsbach PH, Gu K, Franceschi RT, Rutherford RB -2000-. Gene therapy-directed osteogenesis: BMP-7-transduced human fibroblasts form bone in vivo. Hum Gene Ther11:1201–1210.
16.
Lieberman JR, Le LQ, Wu L, Finerman GA, Berk A, Witte ON, et al. -1998-. Regional gene therapy with a BMP-2-producing murine stromal cell line induces heterotopic and orthotopic bone formation in rodents. J Orthop Res16:330–339.
17.
Lieberman JR, Daluiski A, Stevenson S, Wu L, McAllister P, Lee YP, et al. -1999-. The effect of regional gene therapy with bone morphogenetic protein-2-producing bone-marrow cells on the repair of segmental femoral defects in rats. J Bone Joint Surg Am81:905–917.
18.
Manolagas SC, Burton DW, Deftos LJ -1981-. 1,25-Dihydroxyvitamin D3 stimulates the alkaline phosphatase activity of osteoblast-like cells. J Biol Chem256:7115–7117.
19.
Nakamura K, Shirai T, Morishita S, Uchida S, Saeki-Miura K, Makishima F -1999-. p38 mitogen-activated protein kinase functionally contributes to chondrogenesis induced by growth/differentiation factor-5 in ATDC5 cells. Exp Cell Res250:351–363.
20.
Nishitoh H, Ichijo H, Kimura M, Matsumoto T, Makishima F, Yamaguchi A, et al. -1996-. Identification of type I and type II serine/threonine kinase receptors for growth/differentiation factor-5. J Biol Chem271:21345–21352.
21.
Nohe A, Keating E, Knaus P, Petersen NO -2004-. Signal transduction of bone morphogenetic protein receptors. Cell Signal16:291–299.
22.
Peng H, Wright V, Usas A, Gearhart B, Shen HC, Cummins J, et al. -2002-. Synergistic enhancement of bone formation and healing by stem cell-expressed VEGF and bone morphogenetic protein-4. J Clin Invest110:751–759.
23.
Phimphilai M, Zhao Z, Boules H, Roca H, Franceschi RT -2006-. BMP signaling is required for RUNX2-dependent induction of the osteoblast phenotype. J Bone Miner Res21:637–646.
24.
Riew KD, Wright NM, Cheng S, Avioli LV, Lou J -1998-. Induction of bone formation using a recombinant adenoviral vector carrying the human BMP-2 gene in a rabbit spinal fusion model. Calcif Tissue Int63:357–360.
25.
Rutherford RB, Moalli M, Franceschi RT, Wang D, Gu K, Krebsbach PH -2002-. Bone morphogenetic protein-transduced human fibroblasts convert to osteoblasts and form bone in vivo. Tissue Eng8:441–452.
26.
ten Dijke P, Yamashita H, Sampath TK, Reddi AH, Estevez M, Riddle DL, et al. -1994-. Identification of type I receptors for osteogenic protein-1 and bone morphogenetic protein-4. J Biol Chem269:16985–16988.
27.
Winn SR, Hu Y, Sfeir C, Hollinger JO -2000-. Gene therapy approaches for modulating bone regeneration. Adv Drug Deliv Rev42:121–138.
28.
Yamashita H, ten Dijke P, Huylebroeck D, Sampath TK, Andries M, Smith JC, et al. -1995-. Osteogenic protein-1 binds to activin type II receptors and induces certain activin-like effects. J Cell Biol130:217–226.
29.
Zhao M, Zhao Z, Koh JT, Jin T, Franceschi RT -2005-. Combinatorial gene therapy for bone regeneration: cooperative interactions between adenovirus vectors expressing bone morphogenetic proteins 2, 4, and 7. J Cell Biochem95:1–16.
30.
Zhao Z, Zhao M, Xiao G, Franceschi RT -2005-. Gene transfer of the Runx2 transcription factor enhances osteogenic activity of bone marrow stromal cells in vitro and in vivo. Mol Ther12:247–253.
31.
Zhao Z, Wang Z, Ge C, Krebsbach P, Franceschi RT -2007-. Healing cranial defects with AdRunx2-transduced marrow stromal cells. J Dent Res86:1207–1211.
32.
Zhu W, Rawlins BA, Boachie-Adjei O, Myers ER, Arimizu J, Choi E, et al. -2004-. Combined bone morphogenetic protein-2 and -7 gene transfer enhances osteoblastic differentiation and spine fusion in a rodent model. J Bone Miner Res19:2021–2032.
33.
Zhu W, Kim J, Cheng C, Rawlins BA, Boachie-Adjei O, Crystal RG, et al. -2006-. Noggin regulation of bone morphogenetic protein -BMP- 2/7 heterodimer activity in vitro. Bone39:61–71.
