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. Author manuscript; available in PMC: 2015 Mar 10.
Published in final edited form as: Stem Cells. 2011 Aug;29(8):1241–1255. doi: 10.1002/stem.670

Figure 6.

Figure 6

Dura mater (DM) enhances bone morphogenetic protein (BMP) and osteogenic gene expression within mouse calvarial defects and on overlying human adipose-derived stromal cell (hASCs). (A): Histological analysis of the defect center postoperative day 5 with or without DM intact and without scaffolds or hASCs. On the left, an image of the defect site in the right parietal bone (pb) and surrounding calvaria is shown for orientation purposes. Second column: Bmp-2 expression by in situ hybridization (ISH). Third column: Runt-related protein-2 (RUNX-2) expression by ISH. Fourth column: Osteocalcin (Ocn) expression by ISH. Immunohistochemistry analysis of Ocn (fifth column) and phosphorylated SMAD-1 (sixth column). Groups compared from top to bottom include (a) calvarial defect with the DM left intact and (b) calvarial defect with the DM removed. (B): Calvarial defect with the DM intact or removed and with a hASC seeded scaffold placed in the defect. Stains are the same as described in (A). (C): Quantitative real-time polymerase chain reaction analysis of RNA derived from cranial defects at 5-day postoperatively, specific for mouse genes. A significant reduction of mAlp mRunx-2 mCol1a1 mBmp-2 mBmp-4, and mBmpr1b was observed upon DM removal; n = 5 animals per group; *, p < .05. A paired Student’s t test was performed to assess significance. (D): Histological analysis of the defect center postoperative day 5 with DM intact or with DM removed treated with a scaffold with hASCs. On the left, an image of the defect site in the right pb and surrounding calvaria is shown for orientation purposes. Second column: Bmp-2 expression by ISH specific for mouse. (E): Histological analysis of the defect edge postoperative day 5 with DM intact or with DM removed. On the left, an image of the defect site in the right pb and surrounding calvaria is shown for orientation purposes. Second column: Bmp-2 expression by ISH. Third column: Runx-2 expression by ISH. Fourth column: Ocn expression by ISH. Abbreviations: ASC, adipose-derived stromal cell; BMP-2, bone morphogenetic protein; cb, contralateral (uninjured) bone; OCN, osteocalcin; pb, parietal bone; pSMAD, phosphorylated SMAD; RUNX-2, Runt-related protein-2; tb, temporal bone.