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. 2022 Apr 29:9:rbac027.
doi: 10.1093/rb/rbac027. eCollection 2022.

Mg(OH)2 nanosheets on Ti with immunomodulatory function for orthopedic applications

Yue He  1   2 , Mengyu Yao  2 , Jielong Zhou  2 , Juning Xie  1   2 , Changxiang Liang  2 , Dong Yin  2 , Shuaihao Huang  2 , Yu Zhang  1   2 , Feng Peng  2 , Shi Cheng  2
Affiliations

Mg(OH)2 nanosheets on Ti with immunomodulatory function for orthopedic applications

Yue He et al. Regen Biomater. .

Abstract

Macrophages play a vital role for guiding the fate of osteogenesis- related cells. It is well known that nano-topography and bioactive ions can directly enhance osteogenic behavior. However, the effects of nano-structure combined with bioactive ions release on macrophage polarization and the following osteogenesis and angiogenesis are rarely reported. Herein, Mg(OH)2 films with nano-sheet structures were constructed on the surface of Ti using hydrothermal treatment. The film presented nano-sheet topography and sustained release of Mg ions. The results of in vitro culture of bone marrow-derived macrophages (BMDMs), including PCR, western blot and flow cytometry suggested that the nano-Mg(OH)2 films were more favorable for macrophages polarizing to tissue healing M2 phenotype. Moreover, air-pouch model confirmed that the nano-Mg(OH)2 film coated Ti would induce milder inflammation and thinner fibrous layer in vivo, compared with untreated Ti. Furthermore, macrophages-conditioned culture mediums were collected from nano-Mg(OH)2 coated Ti group was superior for the osteogenic behaviors of mice bone marrow stem cells and the angiogenic behaviors of human umbilical vein endothelial cells. With harmonious early inflammatory response and subsequently improved osteogenesis and angiogenesis, the nano-Mg(OH)2 coated Ti is promising for orthopedic applications.

Keywords: immunomodulation; nano-magnesium hydroxide; osteogenesis; titanium.

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Figures

None
Graphical abstract
Figure 1.
Figure 1.
Surface morphology (a) and water contact angle (b) of Ti, HT-1# and HT-2#. Accumulative release of Mg ions from HT-1# and HT-2# (c)
Figure 2.
Figure 2.
Live/dead staining (a) and cell proliferation (b) of BMDMs after cultured on Ti, HT-1# and HT-2# for various time. Flow cytometry analyses of apoptosis marker on macrophages (c) and corresponding quantitative result (d)
Figure 3.
Figure 3.
Adhesion of BMDMs after cultured on Ti, HT-1# and HT-2# surfaces for 12 h
Figure 4.
Figure 4.
Relative mRNA expression of M1 (TNF-α, IL-6, CCL-3, IL-1) and M2 (arg-1, CD163, IL-10) related genes in BMDM after cultured on various samples (a). Expression of CD206 (M2 marker) and iNOS (M1 marker) proteins in BMDMs after cultured on various samples for 3 days (b). Mean fluorescence intensity of CD86 (M1 marker) and CD206 (M2 marker) in BMDMs determined by flow cytometry (c)
Figure 5.
Figure 5.
Immunofluorescence images of iNOS and CD206 in tissues adjacent to various samples (a). H&E staining of the tissues adjacent to various samples (b). The blue dotted lines indicate the fibrous layers
Figure 6.
Figure 6.
Scheme of experiment process (a). ALP activity (b) and ECM mineralization (c) analysis of BMSCs cultured. Relative mRNA expression of osteogenesis-related genes in BMSCs (d). Expression of ALP and RUNX2 proteins in BMSCs and corresponding quantitative analysis (e)
Figure 7.
Figure 7.
Scheme of experiment process (a). Tube formation ability of HUVECs after cultured in MCM for 12 h (b). The calculated tube lengths of the formed vessel (c). Relative mRNA expression of angiogenesis-related genes in HUVECs after cultured in MCM for 3 days (d)
Figure 8.
Figure 8.
Methamphetamine blue staining of the obtained femur bone after implantation for 8 weeks
See this image and copyright information in PMC

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