Protein adhesion and osteoblast differentiation on porous titanium with different porosity and pore size

doi:10.3877/cma.j.issn.1674-1366.2015.03.003

Abstract

Abstract:

Objective

To evaluate the effect of porosity and pore size of the porous titanium on the protein adhesion and osteoblast differentiation from the perspective of molecular biology, thus to provide theoretical evidence for clinical application of porous titanium and to guide the optimal designer of porous titanium.

Methods

Through controlling particle sizes (Ⅰ: 0~200 μm, Ⅱ: 200~400 μm, Ⅲ: 400~600 μm) and weight ratios (A: 20%, B: 30%) of NH₄HCO₃, six groups of porous titanium samples with different porosities and pore sizes were fabricated by powder metallurgy process, dense titanium were made as control group, 10 specimens per group. The adhesive proteins were tested with protein quantification kit to evaluate the protein adsorption capacity of porous titanium. The gene expression of Runt transcription factor 2 (Runx2) , alkaline phosphatase (ALP) , and osteocalcin (OCN) of MC3T3-E1 cells were tested by real-time fluorescence quantitative polymerase chain reaction (PCR) .

Results

All porous titanium groups showed higher protein adsorption than control group (P < 0.05) . The higher the mean porosity is and the smaller the mean pore size is, the more serum proteins adhere to compared with the other porous titanium groups (P < 0.05) . The porosity and pore size have interaction effects on differentiation of osteoblasts. Osteoblasts only expressed higher Runx2 gene on group AⅠ than control group at day 7 (P < 0.05) . While cells on control group expressed significantly higher levels of transcript for Runx2 than the cells on all porous Ti disks at day 14 (P < 0.05) . The gene expression of ALP is significantly higher compared with control group in all porous Ti disks on 7 and 14 day (P < 0.05) , BⅢ at day 7 and AⅠ, AⅢ at day 14 is excluded. For gene expression of OCN, on group AⅠ and BⅠ are higher than on dense Ti at day 7 (P < 0.05) ; on group BⅠ is higher than on dense Ti at day 14 (P < 0.05) .

Conclusions

All porous titanium groups shows higher protein adsorption than control group. The porous titanium with mean porosity of 53.3% and mean pore size of 191.6 μm facilitates the osteoblastic differentiation of mouse osteoblast precursor cells MC3T3-E1.

Key words: Titanium, Porosity, Pore size, Protein adhesion, Osteoblast

Shuai Liu, Xiangxia Li, Yue Yang, Ke Zhao. Protein adhesion and osteoblast differentiation on porous titanium with different porosity and pore size[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2015, 09(03): 193-199.

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Figures/Tables 6

References 19

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组别	平均孔隙率（%）	平均孔隙尺寸（μm）
AⅠ组	43.1 ± 0.7	154.8 ± 11.9
AⅡ组	40.9 ± 1.5	295.6 ± 8.5
AⅢ组	44.3 ± 1.1	560.4 ± 25.6
BⅠ组	53.3 ± 1.2	191.6 ± 3.7
BⅡ组	51.7 ± 2.7	303.8 ± 8.2
BⅢ组	49.9 ± 3.9	583.1 ± 21.7
C组	0	0

组别	平均孔隙率（%）	平均孔隙尺寸（μm）
AⅠ组	43.1 ± 0.7	154.8 ± 11.9
AⅡ组	40.9 ± 1.5	295.6 ± 8.5
AⅢ组	44.3 ± 1.1	560.4 ± 25.6
BⅠ组	53.3 ± 1.2	191.6 ± 3.7
BⅡ组	51.7 ± 2.7	303.8 ± 8.2
BⅢ组	49.9 ± 3.9	583.1 ± 21.7
C组	0	0

基因	引物序列
Runx2	正向：5 ′ -GCCGGGAATGATGAGAACTT-3 ′
?	反向：5 ′ -GGACCGTCCACTGTCACTTA-3 ′
ALP	正向：5 ′ -AACCCAGACACAAGCATTCG-3 ′
?	反向：5 ′ -GCCTTTGAGGTTTTTGGTCT-3 ′
OCN	正向：5 ′ -TTCTGCTCACTCTGCTGACG-3 ′
?	反向：5 ′ -ACCACTCCAGCACAACTCCA-3 ′
β-actin	正向：5 ′ -GCTCTTTTCCAGCCTTCCTA-3 ′
?	反向：5 ′ -GTGCTAGGAGCCAGAGCAGA-3 ′

基因	引物序列
Runx2	正向：5 ′ -GCCGGGAATGATGAGAACTT-3 ′
?	反向：5 ′ -GGACCGTCCACTGTCACTTA-3 ′
ALP	正向：5 ′ -AACCCAGACACAAGCATTCG-3 ′
?	反向：5 ′ -GCCTTTGAGGTTTTTGGTCT-3 ′
OCN	正向：5 ′ -TTCTGCTCACTCTGCTGACG-3 ′
?	反向：5 ′ -ACCACTCCAGCACAACTCCA-3 ′
β-actin	正向：5 ′ -GCTCTTTTCCAGCCTTCCTA-3 ′
?	反向：5 ′ -GTGCTAGGAGCCAGAGCAGA-3 ′

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