In vitro growth and differentiation of osteoblast-like cells on hydroxyapatite ceramic granule calcified from red algae.
OBJECTIVE
The purpose of this study was to analyze the interaction between osteoblast-like cells isolated from mandibular bone and hydroxyapatite ceramic bone substitute obtained from calcified red algae to assess the growth and differentiation of adherent cells on this biomaterial.


METHODS
The macroporous ceramic material C GRAFT/Algipore (The Clinician's Preference LLC, Golden, CO) is composed of 100% hydroxyapatite and possesses specific mechanical and physiochemical properties. Osteoblast-like cells were seeded on 200 mg of biomaterial and cultured for 6 and 21 days under osteogenic differentiation conditions. Specific alkaline phosphatase activity, DNA, and protein content of the proliferating cells were analyzed. The morphology of the cells in contact with the biomaterial was examined by scanning electron microscopy. The osteoblastic phenotype of the cells was confirmed by analysis of the expression of bone-specific genes (osteocalcin, osteopontin and collagen type I) by semi-quantitative reverse transcriptase polymerase chain reaction.


RESULTS
The DNA and protein content increased over the culture period. Scanning electron microscopy showed cells spreading on the surface of the biomaterials, covering the macropores, and colonizing the depth of the particles. The analysis of the expression patterns of bone-related genes confirmed the osteoblastic phenotype of the cultured cells.


CONCLUSIONS
The results of this study showed that hydroxyapatite ceramic bone substitute obtained from calcified red algae support the proliferation and differentiation of human osteoblast-like cells on its surface in vitro and might be suitable for use as scaffolds in tissue engineering strategies in vivo.
Turhani
Dritan
D
Medical University of Vienna, Hospital of Cranio-Maxillofacial and Oral Surgery, Austria. dritan.turhani@meduniwien.ac.at
Cvikl
Barbara
B
Watzinger
Elisabeth
E
Weissenböck
Martina
M
Yerit
Kaan
K
Thurnher
Dietmar
D
Lauer
Günter
G
Ewers
Rolf
R
D018786:Bone Substitutes; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D002516:Ceramics; D024042:Collagen Type I / Q000096:biosynthesis; D017886:Durapatite; D006801:Humans; D008334:Mandible / Q000166:cytology; D008422:Materials Testing; D008855:Microscopy, Electron, Scanning; D010006:Osteoblasts / Q000166:cytology* / Q000378:metabolism; D015675:Osteocalcin / Q000096:biosynthesis; D053495:Osteopontin; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000461:Rhodophyta; D012795:Sialoglycoproteins / Q000096:biosynthesis; D023822:Tissue Engineering
C483718:Algipore; D018786:Bone Substitutes; D024042:Collagen Type I; D012333:RNA, Messenger; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; D015675:Osteocalcin; D053495:Osteopontin; D017886:Durapatite