The influence of different titanium topographies on the biological functions of mesenchymal stem cells

doi:10.3877/cma.j.issn.1674-1366.2017.01.005

Abstract

Abstract:

Objective

To study the influence of different titanium topographies on the adhesion, proliferation and differentiation of mesenchymal stem cells (MSCs) .

Methods

All the titanium discs were divided into three groups and accepted the following treatments: machined smooth Ti (MP) , sandblasted and acid etched Ti (SLA) , TiO₂ nanotubes (TNT) . The topography of Ti discs was observed by a scanning electron microscope (SEM) . The surface roughness of samples was determined using a laser scanning confocal microscope. The contact angles were measured using a contact angle analyzer. The morphology of MSC on different titanium surfaces was evaluated by immunofluorescence staining. Cell early adhesion was evaluated by counting the number of cells on different titanium surfaces. The cell counting kit-8 (CCK-8) assay was used to evaluate the proliferation activity. Alkaline phosphatase (ALP) activity was measured using an ALP kit. The level of significance was determined by One-Way ANOVA followed by LSD-t test for a multiple comparison procedure.

Results

Evenly distributed nanotubes were observed in TNT with a diameter of about 80 nm, while a hierarchical structure with pores and holes was found in SLA. The roughness of SLA (Sa= 1.39 μm, Sq= 1.75 μm) was the highest, followed by TNT (Sa= 1.15 μm, Sq= 1.48 μm) and MP (Sa= 0.87 μm, Sq= 1.14 μm) (F_Sa= 28.54, F_Sq= 24.70, P<0.01) . The contact angle of TNT (8.11°) was the lowest among the three groups (F= 16.32, P<0.01) , indicating the highest hydrophilicity. With the immunofluorescence staining, cells were observed as cuboidal on MP, polygon-like on SLA and spindle-like on TNT. At the first 30 min of culture, more cell attachment was found on TNT and SLA (132.45/field, 176.90/field) than that on MP (64.80/field) (F=12.05, P<0.01) , while there was no significant difference among groups at 60 min of culture. According to the result of CCK-8 assay, there was no significant difference in the proliferation of cells among groups on day 1 and 7; whereas on day 3 and 5, the proliferation level of cells on MP was the highest, followed by TNT and SLA (F_{3 d}= 175.44, P_{3 d}<0.01; F_{5 d}= 6.329, P_{5 d}= 0.02) . A significantly higher ALP activity was detected in TNT (ALP_{7 d}= 156.34 U/gprot, ALP_{14 d}= 153.48 U/gprot) , compared with MP (ALP_{7 d}= 68.21 U/gprot, ALP_{14 d}= 102.73 U/gprot) and SLA (ALP_{7 d}= 65.30 U/gprot, ALP_{14 d}= 86.53 U/gprot) (F_{7 d}= 4.93, P_{7 d}= 0.04; F_{14 d}= 6.49, P_{14 d}= 0.02) .

Conclusion

The surface of TNT presents to be highly hydrophilic and is more favorable for the adhesion, proliferation and differentiation of MSC compared with SLA.

Key words: Titanium, Mesenchymal stem cells, Biological functions, Topography

Jingyan Huang, Anxun Wang, Qin Huang, Jialing Wu, Jie Chen, Yan Wang. The influence of different titanium topographies on the biological functions of mesenchymal stem cells[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2017, 11(01): 25-30.

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References 18

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组别	试件数	Sa值（μm）	Sq值（μm）	表面接触角（°）
MP组	5	0.87 ± 0.05^a	1.14 ± 0.06^a	59.34 ± 13.08^a
TNT组	5	1.15 ± 0.07^b	1.48 ± 0.82^b	8.11 ± 3.61^b
SLA组	5	1.39 ± 0.17^c	1.75 ± 0.21^c	73.66 ± 37.41^a
F值	?	28.54	24.70	16.32
P值	?	0.00	0.00	0.00

组别	试件数	Sa值（μm）	Sq值（μm）	表面接触角（°）
MP组	5	0.87 ± 0.05^a	1.14 ± 0.06^a	59.34 ± 13.08^a
TNT组	5	1.15 ± 0.07^b	1.48 ± 0.82^b	8.11 ± 3.61^b
SLA组	5	1.39 ± 0.17^c	1.75 ± 0.21^c	73.66 ± 37.41^a
F值	?	28.54	24.70	16.32
P值	?	0.00	0.00	0.00

组别	试件数	细胞黏附数（个/视野）
组别	试件数	30 min	60 min
MP组	4	64.80 ± 35.06^a	195.85 ± 65.49
TNT组	4	132.45 ± 37.60^b	241.55 ± 54.33
SLA组	4	176.90 ± 23.01^b	252.20 ± 30.50
F值	?	12.05	1.32
P值	?	0.00	0.32

组别	试件数	细胞黏附数（个/视野）
组别	试件数	30 min	60 min
MP组	4	64.80 ± 35.06^a	195.85 ± 65.49
TNT组	4	132.45 ± 37.60^b	241.55 ± 54.33
SLA组	4	176.90 ± 23.01^b	252.20 ± 30.50
F值	?	12.05	1.32
P值	?	0.00	0.32

组别	试件数	A ₄₅₀
组别	试件数	1d	3d	5d	7d
MP组	4	0.54 ± 0.09	1.88 ± 0.06^a	2.18 ± 0.20^a	2.31 ± 0.12
TNT组	4	0.47 ± 0.04	0.97 ± 0.06^b	1.79 ± 0.43^ab	2.04 ± 0.28
SLA组	4	0.45 ± 0.09	0.70 ± 0.14^c	1.43 ± 0.19^b	1.83 ± 0.26
F值	?	1.72	175.44	6.33	3.00
P值	?	0.23	0.00	0.02	0.13

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