A comparative analysis of phenotype expression in human osteoblasts from heterotopic ossification and normal bone.
OBJECTIVE
Heterotopic ossification (HO) is a pathological bone formation process in which ectopic bone is formed in soft tissue. The formation of bone depends on the expression of the osteoblast phenotype. Earlier studies have shown conflicting results on the expression of phenotype markers of cells originating from HO and normal bone. The hypothesis of the present study is that cells from HO show an altered expression of osteoblast-specific phenotype markers compared to normal osteoblasts. The aims of the study were to further characterize the expression of osteoblast phenotypemarkers and to provide a comparison with other study results.


METHODS
Using an in vitro technique, reverse transcription polymerase chain reaction (RT-PCR), real-time PCR and immunohistochemistry, we compared the phenotype gene expression (type I collagen, alkaline phosphatase, Cbfa-1, osteocalcin) of osteoblasts from resected HO and normal bone (iliac crest).


RESULTS
Cells from HO expressed the osteoblast phenotype (type I collagen, alkaline phosphatase) but were characterized by a depleted osteocalcin expression. The expression of Cbfa-1 (osteocalcin transcription gene) showed a large variety in our study. Preoperative radiotherapy had no effect on phenotype expression in cells from HO.


CONCLUSIONS
Our results provide a characterization of cells originating from HO and support the thesis of an impaired osteoblast differentiation underlying the formation of HO. The transcription axis from Cbfa-1 to osteocalcin could be involved in the pathogenesis of HO.
Handschin
Alexander E
AE
Department of Surgery, Research Division, University Hospital of Zurich, Raemistrasse 100, 8091, Zurich, Switzerland. alexander.handschin@usz.ch
Egermann
Marcus
M
Wedler
Volker
V
Trentz
Otmar
O
Hemmi
Sonja
S
Trentz
Omana A
OA
D000328:Adult; D000469:Alkaline Phosphatase / Q000235:genetics; D016025:Bone Transplantation; D001842:Bone and Bones / Q000378:metabolism / Q000473:pathology; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D005260:Female; D005593:Fracture Fixation, Internal; D015870:Gene Expression / Q000502:physiology; D005819:Genetic Markers / Q000235:genetics*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009999:Ossification, Heterotopic / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D010006:Osteoblasts / Q000378:metabolism* / Q000473:pathology; D015675:Osteocalcin / Q000235:genetics; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D024042:Collagen Type I; D050659:Core Binding Factor Alpha 1 Subunit; D005819:Genetic Markers; D012333:RNA, Messenger; D015675:Osteocalcin; D000469:Alkaline Phosphatase