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. 2020 Jul 24:2020:2327034.
doi: 10.1155/2020/2327034. eCollection 2020.

The Effects of Titanium Surfaces Modified with an Antimicrobial Peptide GL13K by Silanization on Polarization, Anti-Inflammatory, and Proinflammatory Properties of Macrophages

Xuxi Chen  1 , Lin Zhou  2 , Dong Wu  3 , Wenxiu Huang  4 , Yanjun Lin  5 , Bowei Zhou  6 , Jiang Chen  7
Affiliations

The Effects of Titanium Surfaces Modified with an Antimicrobial Peptide GL13K by Silanization on Polarization, Anti-Inflammatory, and Proinflammatory Properties of Macrophages

Xuxi Chen et al. Biomed Res Int. .

Abstract

The polarization of macrophages and its anti-inflammatory and proinflammatory properties play a significant role in host response after implant placement to determine the outcome of osseointegration and long-term survival. In the previous study, we immobilized an antimicrobial peptide, GL13K, onto titanium surfaces to provide immune regulation property. In the herein presented study, we aimed at investigating whether GL13K immobilized titanium surface could improve osteogenesis and reduce the inflammatory reaction around the biomaterials by altering macrophage response. We evaluated the cell proliferation of the different phenotypes of macrophages seeded in GL13K-coated titanium surface, which indicated an inhibition of M1 macrophages and a good cytocompatibility to M2 macrophages. Then, we measured the inflammatory and anti-inflammatory activity of the M1 and M2 macrophages seeded on the GL13K-coated titanium surfaces. The results of the enzyme-linked immunosorbent assay and quantitative reverse transcription-polymerase chain reaction showed that the group with the GL13K modified surface had a downregulation in the expression level of the tumor necrosis factor-α and interleukin-1β in M1 macrophages and an upregulation of IL-10 and transforming growth factor-β3 (TGF-β3) levels in M2 macrophages. This study demonstrated that the GL13K modified titanium surfaces can regulate macrophages' polarization and the expression of inflammatory and anti-inflammatory effects, reducing the effects of the inflammatory process, which may promote the process of bone regeneration and osseointegration.

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Conflict of interest statement

The authors declare that they have no conflicts of interest.

Figures

Figure 1
Figure 1
FACS results of RAW264.7 cells cultured on the titanium surface, the silanized titanium surface, and the GL13K-coated surface for 24 h, 72 h, and 7 d. After being seeded onto GL13K-coated titanium surfaces, the mean expression level of CD206 was increased in comparison to that in the titanium groups, while the mean expression level of CD11c was reduced in the cells cultured on the GL13K-coated titanium compared with the titanium group.
Figure 2
Figure 2
Cell proliferation of M1 macrophages and M2 macrophages after culturing on the titanium surface, the silanized titanium surface, and the GL13K-coated surface for 24, 48, and 72 h. Error bars represent mean ± SD for n = 5. A statistically significant difference exists between GL13K-coated Ti groups and Ti groups at 48 h (p < 0.001) and 72 h (p < 0.05).
Figure 3
Figure 3
ELISA analysis of the secretion of IL-1β and TNF-α in macrophages with M1 polarization and IL-10 and IL-1ra in macrophages with M2 polarization cultured on the titanium surface, the silanized titanium surface, and the GL13K-coated surface at 24 h. Error bars represent mean ± SD for n = 5. The downregulation of the secretion level of cytokines TNF-α (p < 0.001) and the upregulation of cytokines IL-10 (p = 0.002) were detected in the GL13K immobilized titanium group.
Figure 4
Figure 4
Quantitative analysis of the secretion of TNF-α, IL-6, and IL-1β of macrophages with M1 polarization and IL-10, IL-1ra, TGF-β1, and TGF-β3 of macrophages with M2 polarization after culturing on the titanium surface, the silanized titanium surface, and the GL13K-coated surface for 3 days. Error bars represent mean ± SD for n = 5. A downregulation of the relative mRNA expression of TNF-α (p = 0.001) and IL-1β (p < 0.001) was detected in the groups of GL13K immobilized titanium, while the expression of IL-10 (p < 0.001) and TGF-β3 (p = 0.003) levels were upregulated in these groups.
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