The effect of globin scaffold on osteoblast adhesion and phenotype expression in vitro.
OBJECTIVE
Different synthetic and natural biomaterials have been used in bone tissue regeneration. However, several limitations are associated with the use of synthetic as well as allogenous or xenogenous natural materials. This study evaluated, in an in vitro model, the behavior of rat osteoblastic cells cultured on a human globin scaffold.


METHODS
Rat osteoblastic cells were isolated from the calvaria of 21-day-old fetal Sprague-Dawley rats. They were then grown in the presence of globin. Real-time polymerase chain reaction (RT-PCR) was performed to study the expression of cyclin D1, integrin Β1, Msx2, Dlx5, Runx2, and osteocalcin on days 1, 5, and 9. Moreover, alkaline phosphatase activity was measured on days 1, 3, 5, and 7. Alizarin red staining was performed on day 9 to observe calcium deposition.


RESULTS
Cells were able to adhere, proliferate, and differentiate on globin scaffolds. Moreover, RT-PCR showed that globin may stimulate some key genes of osteoblastic differentiation (Runx2, osteocalcin, Dlx5). Globin had an inhibitory effect on alkaline phosphatase activity. Calcium deposits were seen after 9 days of culture.


CONCLUSIONS
These results indicate that purified human globin might be a suitable scaffold for bone tissue regeneration.
INSERM, Laboartoire de Physiopathologie Orale Moléculaire, Centre de Recherche des Cordeliers, Université Pierre et Marie Curie-Paris 6, Université Paris Descartes, Paris, France. ahmad.hamdan@ju.edu.jo
Ahmad A
AA
Hamdan
Sabine
S
Loty
Juliane
J
Isaac
Jean-Louis
JL
Tayot
Philippe
P
Bouchard
Ameen
A
Khraisat
Ariane
A
Bedral
Jean-Michel
JM
Sautier
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001861:Bone Regeneration; D001842:Bone and Bones / Q000737:chemistry / Q000378:metabolism; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000378:metabolism; D019938:Cyclin D1 / Q000378:metabolism; D005914:Globins / Q000494:pharmacology*; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D015675:Osteocalcin / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D054457:Tissue Scaffolds; D014157:Transcription Factors / Q000378:metabolism
D050659:Core Binding Factor Alpha 1 Subunit; C088334:Distal-less homeobox proteins; D018398:Homeodomain Proteins; D019012:Integrin beta1; C084119:MSX2 protein; C493717:Runx2 protein, rat; D014157:Transcription Factors; D015675:Osteocalcin; D019938:Cyclin D1; D005914:Globins; D000469:Alkaline Phosphatase