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. Author manuscript; available in PMC: 2009 Mar 9.
Published in final edited form as: Oral Dis. 2008 Jul;14(5):428–434. doi: 10.1111/j.1601-0825.2007.01396.x

Figure 2.

Figure 2

Characterization of SHED-mediated bone formation. After 6 months of transplantation, SHED were capable of maintaining bone structure (B) on the surfaces of hydroxyapatite/tricalcium phosphate (HA) along with connective tissue (CT, a). Same field of polarized picture showed dense collagen fibers (b). In contrast, bone marrow mesenchymal stem cells maintained both bone (B) and bone marrow elements (BM) after 6 months posttransplantation (c). Same field of polarized microscopy view showed dense collagen fibers (d). In situ hybridization studies showed the murine-specific pf1 DNA probe reacting with recipient osteoblasts and osteocytes associated with the new bone formation (B, black arrows in e). Mouse tissue (MT) reacted positive for pf1 probe in SHED transplant (open arrows in e). Human-specific alu in situ hybridization showed that SHED (black arrows in f) were associated with bone formation (B). Mouse tissue (MT) was negative for alu in situ hybridization. Immunohistochemical staining showed that SHED generated bone (B) and differentiated into osteocytes that were positive for anti-human-specific mitochondria antibody staining (open arrows in g). Mouse tissue (MT in g) and preimmunoserum control (h) were negative for anti-human-specific mitochondria antibody staining. Single colony-derived SHED were also capable of forming bone (B) on the surface of hydroxyapatite/tricalcium phosphate (HA) to repair critical size of calvarial defects (black line in i) in immunocompromised mice (i, j) same as seen in mixed population of SHED. CB indicates preexisting calvarial bone. The mRNA isolated from two different SHED transplants (T1 and T2) was applied for RT-PCR analysis (k). Both human (h) and mouse (m) BSP and OSC were positively detected on both transplants, suggesting that both human and mouse osteogenic cells involved in the bone formation in the SHED transplants. The mRNA extracted from human (H) and mouse (M) intact bones were used as positive and negative controls for RT-PCR amplification. GAPDH was used for internal control