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Healing Cranial Defects with AdRunx2-transduced Marrow Stromal Cells

¹ Program in Oral Health Sciences,
² Department of Periodontics and Oral Medicine, and
³ Department of Biological and Material Sciences, School of Dentistry, University of Michigan, 1011 N. University Ave., Ann Arbor, MI 48109-1078, USA; and
⁴ Department of Biological Chemistry, School of Medicine, University of Michigan, Ann Arbor, MI, USA

View larger version (19K): [in this window] [in a new window]   Figure 1. Induction of osteoblast-related mRNAs in vitro. MSCs were transduced with Ad-LacZ or Ad-Runx2 at a titer of 300 pfu/cell. Cells were harvested at the times indicated. Total RNA was extracted and analyzed for expression of Runx2 (A), ALP (B), BSP (C), and OCN (D) mRNAs, by means of quantitative real-time PCR, as described in MATERIALS & METHODS. All mRNA levels are expressed relative to levels of GAPDH mRNA, which remained constant throughout the experiment. Data are means ± SD of 3 independent samples.

View larger version (20K): [in this window] [in a new window]   Figure 2. In vivo gene expression pattern after MSC implantation. MSCs were transduced with 300 pfu/cell of the indicated adenovirus. Twenty-four hours after transduction, 2 x 106 cells were seeded into gelatin sponges and implanted subcutaneously into immunodeficient mice, as described in MATERIALS & METHODS. Implants were harvested at the times indicated. Total RNA was extracted and analyzed by quantitative real-time PCR (as in Fig. 1). Data are means + SD of 3 independent samples.

View larger version (54K): [in this window] [in a new window]   Figure 3. Repair of critical-sized calvarial defects with Runx2 genetically engineered marrow stromal cells. MSCs were transduced with 300 pfu/cell of the indicated adenovirus. After 24 hrs, 5 x 106 cells were seeded on a gelatin sponge and implanted into 5-mm calvarial defects as described in MATERIALS & METHODS. After 7 wks, implants were harvested and analyzed by x-ray (A), histology (B,C), and micro-CT (D). Panel C shows a higher magnification of the rectangular regions shown in B. Representative 3-D reconstruction images with micro-CT in each group are shown in panel D. Bars = (in panel B) 500 µm and (in panel C) 50 µm.

Journal of Dental Research, Vol. 86, No. 12, 1207-1211 (2007)
DOI: 10.1177/154405910708601213


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