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. 2020 Sep 8:15:6593-6603.
doi: 10.2147/IJN.S255486. eCollection 2020.

A Magnesium-Incorporated Nanoporous Titanium Coating for Rapid Osseointegration

Xiaodong Li #  1   2   3 , Mingyi Wang #  3 , Wenjie Zhang  4 , Yuting Bai  2 , Yuan Liu  3 , Jian Meng  1   2 , Ling Zhang  3
Affiliations
Free PMC article

A Magnesium-Incorporated Nanoporous Titanium Coating for Rapid Osseointegration

Xiaodong Li et al. Int J Nanomedicine. .
Free PMC article

Abstract

Purpose: Micro-arc oxidation (MAO) is a fast and effective method to prepare nanoporous coatings with high biological activity and bonding strength. Simple micro/nano-coatings cannot fully meet the requirements of osteogenesis. To further improve the biological activity of a titanium surface, we successfully added biological magnesium (Mg2+) to a coating by micro-arc oxidation and evaluated the optimal magnesium concentration in the electrolyte, biocompatibility, cell adhesion, proliferation, and osteogenesis in vitro.

Methods: Nanoporous titanium coatings with different concentrations of magnesium were prepared by micro-arc oxidation and characterized by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). The Mg2+ release ability of the magnesium-incorporated nanoporous titanium coatings was determined by inductively coupled plasma emission spectrometry (ICP-OES). The cytotoxicity of the magnesium-incorporated nanoporous titanium coatings was detected with live/dead double-staining tests. A CCK-8 assay was employed to evaluate cell proliferation, and FITC-phalloidin was used to determine the structure of the cytoskeleton by staining β-actin. Alkaline phosphatase (ALP) activity was evaluated by alizarin red S (ARS) staining to determine the effect of the coatings on osteogenic differentiation in vitro. The mRNA expression of osteogenic differentiation-related markers was measured using qRT-PCR.

Results: EDS analyses revealed the successful addition of magnesium to the microporous coatings. The best magnesium concentration of the electrolyte for preparing the new coating was determined. The results showed that the nano-coatings prepared using the electrolyte with 2 g/L magnesium acetate best promoted the adhesion, proliferation, and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs).

Conclusion: These results suggest that the new titanium metal coating with a dual effect of promoting bone morphology and supplying the biological ion Mg2+ can be beneficial for rapid osseointegration.

Keywords: magnesium-incorporated nanoporous coating; micro/arc oxidation; osteoinductivity.

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Conflict of interest statement

The authors report no conflicts of interest in this work.

Figures

Figure 1
Figure 1
(A) SEM images of six sample surfaces. (B) The roughness of the five coatings (C) The frequency distribution of the pore size. (D) The pore size of the five coatings.
Figure 2
Figure 2
The contact angle of the six sample (*p<0.05 compared with Ti).
Figure 3
Figure 3
EDS and elemental analysis of the six sample.
Figure 4
Figure 4
ICP detection of magnesium ion release from various coatings.
Figure 5
Figure 5
(A) Live/dead assay of cells cultured for 24 h on the samples. Scale bar = 100 μm. (B) Statistical results of the live/dead assay. (C) Cell proliferation measured by CCK-8 assay after 1, 3, and 7 days of culture. (D) Fluorescence images of actin (red) and nuclei (blue) in cells cultured on the samples for 6 h. Scale bar = 100 μm. (E) The average area of cell actin was determined by ImageJ (*p<0.05 compared with Ti and MAO; #p<0.05 compared with TI, MAO, M1, M4, and M8).
Figure 6
Figure 6
ALP activity and calcium deposition. (A) After 7 days of culture, BMSCs on the different coatings were stained with ALP. (B) ALP activity was measured was assayed using a quantitative colorimetric assay. (C) After 21 days of culture, BMSCs on the different coatings were stained with ARS. (D) Calcium deposition activity was assayed using a quantitative colorimetric assay (*p<0.05 compared with Ti and MAO; #p<0.05 compared with TI, MAO, M1, M4, and M8).
Figure 7
Figure 7
Expression of osteogenic-related genes was measured by qRT-PCR after incubating on different coatings for 7 days (n=3). (A) Runx2 expression. (B) ALP expression. (C) OCN expression (*p<0.05 compared with Ti and MAO; #p<0.05 compared with TI, MAO, M1, M4, and M8).
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Grants and funding

This work was supported by the National Natural Science Foundation of China (No. 81771127), the Xuzhou Medical Innovation and Technological Breakthrough Team Project (XWCX201604), the Jiangsu Health and Family Planning Commission Project of Scientific Research (H2017080), and the Xuzhou Science and Technology Project (KC17196).

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