Effects of polymer assisted deposition of Ta2O5 coating on the surface of TiO2 nanotubes on improving the biological activity of MC3T3-E1 cells

doi:10.11958/20230648

Abstract

Abstract:

Objective To explore the effect of polymer assisted deposition of tantalum pentoxide (Ta₂O₅) coating on the surface of TiO₂ nanotubes on the biological activity of mouse bone forming precursor cells, MC3T3-E1 cells. Methods Three sets of samples were prepared, including Ta/TiO₂nanotubes (the Ta/NT group), Ta₂O₅/pure titanium (the Ta₂O₅/PT group) and Ta₂O₅/ TiO₂ nanotubes (the Ta₂O₅/NT group). The Ta₂O₅/PT group and the Ta₂O₅/NT group were prepared by polymer-assisted deposition. The three sets of samples were characterized and tested by scanning electron microscopy (SEM) to observe the surface morphology. X-ray photoelectron spectroscopy (XPS) was used to analyze surface element composition, and X-ray diffraction (XRD) was used to detect surface compounds. MC3T3-E1 cells were selected to explore the effect of three groups of samples on cell biological activity. Fluorescence microscope was used to observe the adhesion of cytoskeleton and nucleus. Cell activity was determined by CCK-8 method. Cell alkaline phosphatase (ALP) activity was detected. Calcium deposition was detected by alizarin red staining and semi-quantitative analysis. The expression levels of recombinant human bone morphogenetic protein-2 (BMP-2), ALP, osteocalcin (OCN) and osteopontin (OPN) were detected by real-time PCR. Results SEM images showed that on the surface of the Ta₂O₅/NT group, the TiO₂ nanotube substrate was covered with a sheet-like, banded cross-linked micro-nano Ta₂O₅ coating structure. XRD showed that the TiO₂ nanotubes in the Ta₂O₅/NT group presented a trace of demineralized titanium and rutile crystal structure. Compared with the Ta/NT group, the Ta₂O₅/NT group and the Ta₂O₅/PT group showed higher cell density, larger cytoskeletal mass and filamentous structure, more crosslinking and clusters, higher cytoskeletal tension and higher cell activity. The semi-quantitative analysis of alizarin red staining samples showed that calcium deposition decreased successively in the Ta₂O₅/NT group, the Ta/NT group and the Ta₂O₅/PT group (P<0.01). The ALP activity, calcium deposition and ALP, OCN and OPN mRNA contents on day 7 and 14 were decreased successively in the Ta₂O₅/NT group, the Ta/NT group and the Ta₂O₅/PT group, and the BMP-2 mRNA content on day 14 in the Ta₂O₅/NT group was the highest in the three groups (P<0.05). Conclusion Ta₂O₅/TiO₂ nanotube composite coating can promote the adhesion of MC3T3-E1 cells, the expression of mineralization and osteogenic genes, and enhance the activities of cell and ALP, which is conducive to the play of biological activities.

Key words: Titanium, nanotubes, Tantalum, Tantalum pentoxide coating, polymer-assisted deposition, surface characterization, biological activity

CLC Number:

R783.1

Figures/Tables 7

References 21

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组别	1 d	4 d	7 d
Ta/NT组	0.267±0.015	0.403±0.021	0.633±0.015
Ta₂O₅/PT组	0.307±0.015^a	0.493±0.015^a	0.873±0.021^a
Ta₂O₅/NT组	0.330±0.020^a	0.487±0.015^a	0.860±0.017^a
F	10.654^＊	235.148^＊＊	169.379^＊＊

组别	1 d	4 d	7 d
Ta/NT组	0.267±0.015	0.403±0.021	0.633±0.015
Ta₂O₅/PT组	0.307±0.015^a	0.493±0.015^a	0.873±0.021^a
Ta₂O₅/NT组	0.330±0.020^a	0.487±0.015^a	0.860±0.017^a
F	10.654^＊	235.148^＊＊	169.379^＊＊

组别	7 d	14 d
Ta/NT组	2.993±0.051	2.420±0.072
Ta₂O₅/PT组	2.577±0.132^a	2.133±0.112^a
Ta₂O₅/NT组	3.197±0.083^ab	2.677±0.112^ab
F	33.298^＊＊	21.968^＊＊

组别	7 d	14 d
Ta/NT组	2.993±0.051	2.420±0.072
Ta₂O₅/PT组	2.577±0.132^a	2.133±0.112^a
Ta₂O₅/NT组	3.197±0.083^ab	2.677±0.112^ab
F	33.298^＊＊	21.968^＊＊

组别	ALP		OCN		OPN		BMP-2
组别	7 d	14 d	7 d	14 d	7 d	14 d	7 d	14 d
Ta/NT组	2.326±0.114	3.015±0.210	2.696±0.116	8.307±0.396	1.978±0.082	0.699±0.020	2.089±0.241	5.570±0.692
Ta₂O₅/PT组	1.690±0.108^a	2.590±0.154^a	0.840±0.070^a	1.013±0.121^a	1.003±0.106^a	0.553±0.078^a	0.913±0.060^a	4.353±0.235^a
Ta₂O₅/NT组	2.563±0.091^ab	3.727±0.078^ab	3.063±0.138^ab	10.283±0.286^ab	2.433±0.120^ab	0.810±0.044^ab	1.670±0.111^ab	6.727±0.250^ab
F	55.895^＊＊	40.233^＊＊	324.817^＊＊	847.407^＊＊	148.039^＊＊	17.871^＊＊	43.111^＊＊	21.236^＊＊

Effects of polymer assisted deposition of Ta₂O₅ coating on the surface of TiO₂ nanotubes on improving the biological activity of MC3T3-E1 cells

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基因名称	引物序列（5′→3′）	产物大小（bp）
BMP-2	上游：TGTGGACTTCAGTGATGTG	59
BMP-2	下游：TGGAGTTCAGGTGGTCAG	59
OCN	上游：TTCTGCTCACTCTGCTGACC	43
OCN	下游：GCCGGAGTCTGTTCACTACC	43
ALP	上游：GGCTACTTCTTCGTGGAG	82
ALP	下游：GCGTGAGTGTTCTTGTCT	82
OPN	上游：GCTTGGCTTATGGACTGA	65
OPN	下游：TAGACTCACCGCTCTTCA	65
GAPDH	上游：AGAAGGTGGTGAAGCAGGCATCT	125
GAPDH	下游：CGGCATCGAAGGTGGAAGAGTG	125

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