Fig. 6

Pamidronate restores in vivo mineralized tissue-forming capacity in bilirubin-impaired SHED. a Schemata of SHED cultures and subcutaneous transplantation (Tx) into immunocompromised Balb/c nu/nu mice (nu/nu). SHED were precultured with 0 μM bilirubin (B0), 50 μM bilirubin (B50), and 50 μM bilirubin plus 10 μM pamidronate (B50+PAM) and were subcutaneously transplanted with hydroxyapatite/tricalcium phosphate particles (HA/TCP) in immunocompromised mice. b–d Histological analysis was performed after 4 weeks of the transplantation. Representative transplant images by hematoxylin and eosin staining (HE) were shown (b). De novo mineralized tissue area in SHED transplants was measured as described in the “Methods” section. n = 5 for all groups. Statistical analysis was performed as described in the “Methods” section. Graph bars showed the means ± SD. *P < 0.05. NS, no significance (c). Representative transplant images by immunofluorescence with anti-human CD146 antibody were shown. White-dot circled area: newly formed mineralized area (d). b, d CT, connective tissue; HA, HA/TCP; MM, mineralized tissue. Bar = 100 μm