Effects of TiO2 nanotube layers on RAW 264.7 macrophage behaviour and bone morphogenetic protein-2 expression.
OBJECTIVE
To investigate behaviour and osteogenic cytokine expression of RAW264.7 macrophages grown on TiO2 nanotube layers.


METHODS
The murine macrophage cell line RAW 264.7 was cultured on TiO2 nanotubes of varying diameter; macrophage morphology was examined using scanning electron microscopy. Cell adhesion and viability were assessed with the aid of the MTT method and BMP-2 and TGF-β gene expression were examined by RT-PCR analysis. Levels of BMP-2, TGF-β1 and ICAM-1 proteins secreted into the supernatant were measured by ELISA assay.


RESULTS
Macrophages cultured on nanotube layers had spread out morphology, the largest (120 nm) nanotube layer eliciting an elongation by 24 h. Macrophages adhered significantly less to 120 nm TiO2 nanotubes than to control discs at 4 h after application; after 24 h incubation, macrophages were sufficiently viable (P < 0.05) on 30 and 70 nm nanotube layers. Increasing nanotube diameter led to increased BMP-2 protein secretion and increased BMP-2 mRNA expression.


CONCLUSIONS
These results demonstrate that nanoscale topography of TiO2 nanotube layers can affect macrophage morphology, adhesion, viability and BMP-2 expression. Macrophages grown on layers of large nanotubes had the highest potential to enhance bone formation during bone healing.
Department of Prosthodontics, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai, 200011, China.
S J
SJ
Sun
W Q
WQ
Yu
Y L
YL
Zhang
X Q
XQ
Jiang
F Q
FQ
Zhang
D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D015870:Gene Expression / Q000187:drug effects* / Q000235:genetics; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D051379:Mice; D043942:Nanotubes; D012333:RNA, Messenger / Q000235:genetics; D014025:Titanium / Q000494:pharmacology*; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism
C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D018799:Intercellular Adhesion Molecule-1; C009495:titanium dioxide; D014025:Titanium