Abstract
Objective To investigate the effect of hydrophilicity of titanium with micro/nanotopographical surface on the cell viability and osteogenic differentiation in rat bone marrow mesenchymal stem cells (rBMSCs). Methods Methods of anodic oxidation and sandblast-alkali heat were utilized to establish scale structures. rBMSCs were seeded on these two titanium discs, cell viability was detected at 1, 3, 5 and 7 days by CCK8 assay. Total protein values and alkaline phosphatase (ALP) activity were examined at 7 and 14 days. ALP, collagen-I(COL1) and runt related transcription factor 2 (RUNX2) mRNA expression at 7 days were detected by qRT-PCR. The t test was used to compare water static contact angle, CCK8, total protein concentration and PCR detection index of osteogenic differentiation. Results The titanium surface treated by anodic oxidation was regular ordered nanotube arrays, while that treated by sandblast-alkali heat exhibited a three dimensional mesh nano porous structure, which both groups presented a similar micro-nanotopographical feature. However, the static contact angle of water in anodic oxidation group was significantly enhanced than that of sandblast-alkali heat group (83.3±2.3 vs 47.7±2.0, t=11.54, P<0.001). Compared with anodic oxidation group, rBMSCs on the discs modified by sandblast-alkali heat showed enhanced cell viability at 3, 5 and 7 days (0.66 ±0.03 vs 0.52 ±0.03, 1.15 ±0.06 vs 0.85 ±0.05, 1.58 ±0.07 vs 1.26 ±0.07), and these differences were statistically significant (t=2.962、3.845、3.183, P=0.042、0.018、0.033). After 7 and 14 days, the total protein concentration of rBMSCs in sandblasting alkali heat group was higher than that of anodic oxidation group [(389±45) μg/ml vs (226±32) μg/ml, (1070±59) μg/ml vs (760±65) μg/ml], whose differences were statistically significant (t=3.319、3.518, P=0.029、0.025). Moreover, ALP activity of rBMSCs of sandblasting alkali heat group at 7 days and 14 days was higher than that of anodic oxidation group [(2.11±0.32) U/gprot vs (1.00±0.21) U/gprot, (6.13±0.57) U/gprot vs (3.92±0.51) U/gprot], which showed statistically significant difference (t=2.912、2.976, P=0.043、0.041). Finally, mRNA levels of ALP, COL1 and RUNX2 in rBMSCs of sandblasting alkali heat group is higher than that of anodic oxidation group at 7 days (1.86 ±0.24 vs 1.00 ±0.15; 2.05 ±0.16 vs 1.00 ±0.14; 2.28 ±0.18 vs 1.00 ±0.12), and these difference was statistically significant (t=3.383、5.012、5.710, P=0.028、0.007、0.005). Conclusion Micro-nanotopographical surface of titanium implant in sandblast-alkali heat group promotes cell viability and osteogenic differentiation of rBMSCs, which may be affected by its hydrophilicity. Key words: Rat bone marrow mesenchymal stem cells; Osteogenic differentiation; Micro-nanotopographical surface; Hydrophilicity
Published Version
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