Effect of Bone Morphogenetic Protein-2-Expressing Muscle-Derived Cells on Healing of Critical-Sized Bone Defects in Mice

ArticleinThe Journal of Bone and Joint Surgery 83-A(7):1032-9 · August 2001with16 Reads
Source: PubMed
Abstract
Cells that express bone morphogenetic protein-2 (BMP-2) can now be prepared by transduction with adenovirus containing BMP-2 cDNA. Skeletal muscle tissue contains cells that differentiate into osteoblasts on stimulation with BMP-2. The objectives of this study were to prepare BMP-2-expressing muscle-derived cells by transduction of these cells with an adenovirus containing BMP-2 cDNA and to determine whether the BMP-2-expressing muscle-derived cells would elicit the healing of critical-sized bone defects in mice. Primary cultures of muscle-derived cells from a normal male mouse were transduced with adenovirus encoding the recombinant human BMP-2 gene (adBMP-2). These cells (5 yen 10(5)) were implanted into a 5-mm-diameter critical-sized skull defect in female SCID (severe combined immunodeficiency strain) mice with use of a collagen sponge as a scaffold. Healing in the treatment and control groups was examined grossly and histologically at two and four weeks. Implanted cells were identified in vivo with use of the Y-chromosome-specific fluorescent in situ hybridization (FISH) technique, and their differentiation into osteogenic cells was demonstrated by osteocalcin immunohistochemistry. Skull defects treated with muscle cells that had been genetically engineered to express BMP-2 had >85% closure within two weeks and 95% to 100% closure within four weeks. Control groups in which the defect was not treated (group 1), treated with collagen only (group 2), or treated with collagen and muscle cells without adBMP-2 (group 3) showed at most 30% to 40% closure of the defect by four weeks, and the majority of the skull defects in those groups showed no healing. Analysis of injected cells in group 4, with the Y-chromosome-specific FISH technique showed that the majority of the transplanted cells were located on the surfaces of the newly formed bone, but a small fraction (approximately 5%) was identified within the osteocyte lacunae of the new bone. Implanted cells found in the new bone stained immunohistochemically for osteocalcin, indicating that they had differentiated in vivo into osteogenic cells. This study demonstrates that cells derived from muscle tissue that have been genetically engineered to express BMP-2 elicit the healing of critical-sized skull defects in mice. The cells derived from muscle tissue appear to enhance bone-healing by differentiating into osteoblasts in vivo. Clinical Relevance: Ex vivo gene therapy with muscle-derived cells that have been genetically engineered to express BMP-2 may be used to treat nonhealing bone defects. In addition, muscle-derived cells appear to include stem cells, which are easily obtained with muscle biopsy and could be used in gene therapy to deliver BMP-2.
    • "Data suggested significant repair of surgical defect two weeks after the transplant, attributed to osteogenesis by transplanted MSCs [119]. Recently, Jiang et al., in 2012, employed a different approach in transplanting MSCs to rabbit calvarial defect model, with and without osteogenic induction, by using platelet rich plasma as a scaffold [120]. More lately, another study employing rat calvarial defect model, MSCs (allogeneic) were loaded onto chitosan/alginate/hydroxyapatite scaffold (CAH), with and without BMP-2 impregnation. "
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    • "BMSCs were isolated from bone marrow, even though recent reports indicate that BMSC can be isolated from other sources, such as peripheral blood (Kuznetsov et al., 2001), fat (Zuk et al., 2001), skin (Mizuno and Glowacki, 1996), vasculature (Brighton et al., 1992), and muscle (Lee et al., 2001). BMSCs were isolated as described by Woodbury (Sarvestani et al., 2010). "
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    • "Athymic nude mice injected with BMP9- transduced C2C12 cells into the quadriceps muscles demonstrated significant orthotopic bone formation on both X-ray and histologic evaluation [1, 2, 57]. Several recent studies have suggested that skeletal muscle may harbor multipotent MSCs as well as osteoblastic progenitor cells [28, 33, 74] . When adenovirally-delivered BMP9 (AdBMP9) was directly injected into the quadriceps muscles of athymic nude mice, there was evidence of increased osteoid matrix production and formation of mature lamellar bone compared to BMP2-and 7-treated groups. "
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